KR101179508B1 - Novel 1,6-disubstituted-3-amino-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridin-7-one compounds and preparation thereof - Google Patents

Abstract

The present invention provides a novel 1,6-disubstituted-4,5,6,7-tetrahydro-1 H -pyrazolo [3,4- c ] pyridin-7-one compound and its pharmaceutically acceptable salt compound, The present invention relates to a method for producing the compound, and to a pharmaceutical composition for an anticancer agent comprising the compound as an active ingredient.

Description

1,6-disubstituted-3-amino-4,5,6,7-tetrahydro-1H-pyrazolo [3,4-c] pyridin-7-one compound and method for preparing the compound {Novel 1,6- disubstituted-3-amino-4,5,6,7-tetrahydro-1H-pyrazolo [3,4-c] pyridin-7-one compounds and preparation girls}

The present invention provides a novel 1,6-disubstituted-4,5,6,7-tetrahydro-1 H -pyrazolo [3,4- c ] pyridin-7-one compound and a pharmaceutically acceptable salt compound thereof; It relates to a method for producing the new compound, and an anticancer drug composition comprising the new compound as an active ingredient.

Cancer is the process by which tumor cells caused by abnormal cell growth kill normal tissues and promote uncontrolled cell colonization, spreading to surrounding tissues, eventually other organs of the body, causing much pain and ultimately death to patients. It is a refractory disease. Cancer causes are largely divided into internal factors and external factors. Internal factors include genetic and immunological factors, and external factors include chemicals, radiation, and viral infections. The mechanism by which normal cells are transformed into cancer cells is not yet known, but more than 80% of them are considered to be caused by external factors. Cancer genes are classified into blood and solid cancers due to the imbalance between oncogenes and tumor suppressor genes. Cancer occurs in all parts of the body, including lung cancer, stomach cancer, liver cancer, breast cancer, uterine cancer, esophageal cancer, prostate cancer, colon cancer, and skin cancer.

Cancer is treated by surgery, radiation therapy, and the use of chemotherapy. In particular, as the detection of cancer becomes possible early due to the development of diagnostic functions, the therapeutic effect by chemotherapeutic agents is increasing. Nevertheless, the development of anticancer drugs of the mechanism is still insufficient. Therefore, the use of anticancer agents suitable for the precise pathogenesis of various mechanisms is limited, and the problem of drug resistance remains a big problem to be solved.

Targets of anticancer agents known to date include protein kinase (mainly PKBa (Akt1), PKCb), aurora kinase (ARK1, ARK2, ARK3), cyclin dependent kinase (mainly CDK 2), glycogen synthesis kinase (mainly GSK-3β), Raf Serine / threonine kinase and threonine kinase and envelope cell growth factor receptor (EGFR), human envelope cell growth factor receptor (HER1, HER2, HER3, HER4), stem cell factor receptor (c-Kit), Rearraged during Transfection Receptor (RET), vascular endothelial growth factor (VEGF-1 (Flt1), VEGF-2 (KDR), VEGF-3 (Flt4)), platelet derived growth factor receptor (PDGFR), bcr-abl kinase, src kinase The same tyrosine kinase and the like are known. In addition, carbonic anhydrase, caspases 1, 2, 3, 8, 9, and tubulin are known targets for colon cancer.

The present inventors have newly designed and synthesized 1,6-disubstituted-3-amino-4,5,6,7-tetrahydro- 1H -pyrazolo [3,4- c ] pyridin-7-one compound in various cancers. By confirming the growth inhibitory effect on the cell line was found to be useful as an anticancer agent has been completed the present invention.

The present invention provides a novel 1,6-disubstituted-3-amino-4,5,6,7-tetrahydro-1 H -pyrazolo [3,4- c ] pyridin-7-one compound or pharmaceutically acceptable Its purpose is to provide its salt compounds.

In addition, the present invention, because the novel compounds exhibit a strong growth inhibitory effect on various cancer cell lines, it is another object to provide an anticancer agent and pharmaceutical composition comprising the novel compounds as an active ingredient.

The present invention also provides a method for preparing the 1,6-disubstituted-3-amino-4,5,6,7-tetrahydro- 1H -pyrazolo [3,4- c ] pyridin-7-one compound. There is another purpose to provide.

The present invention is also used for the synthesis of the above-mentioned 1,6-disubstituted-3-amino-4,5,6,7-tetrahydro- 1H -pyrazolo [3,4- c ] pyridin-7-one compound. It is another object to provide novel intermediate compounds.

In order to achieve the above object, the present invention provides a 1,6-disubstituted-3-amino-4,5,6,7-tetrahydro-1 H -pyrazolo [3, 4- c ] pyridin-7-one compound or a pharmaceutically acceptable salt thereof.

In Chemical Formula 1, n represents an integer of 1, 2, or 3, and R ¹ and R ² represent the same as or different from each other, and R- (CH ₂ ) _ℓ- , wherein R represents halo, C ₁ -C ₈ Unsubstituted or substituted with 1 to 3 substituents selected from alkyl, haloC ₁ -C ₈ alkyl, C ₁ -C ₈ alkoxy, amino, C ₁ -C ₈ alkylamino, and di (C ₁ -C ₈ alkyl) amino Phenyl group; Furyl group; Pyridyl groups; Or a benzodioxol group, and 1 represents an integer of 0, 1, 2 or 3.

1,6-disubstituted-3-amino-4,5,6,7-tetrahydro- 1H -pyrazolo [3,4- c ] pyridin-7-one compound represented by Chemical Formula 1 according to the present invention or Pharmaceutically acceptable salts thereof are useful as anticancer agents and auxiliaries because they exhibit excellent inhibitory activity against various cancer cells.

The compound represented by Chemical Formula 1 according to the present invention may have a chiral center. Accordingly, the category of the compound characterized by the present invention includes all isomers, isomer mixtures, or racemic compounds of the compound represented by Chemical Formula 1. In addition, the category of the compound characterized by the present invention includes a radioactive derivative of the compound represented by the formula (1), these radioactive compounds can be usefully used in the field of biological research.

Hereinafter, the substituents used in the definition of the compound represented by Chemical Formula 1 according to the present invention will be described in more detail.

'Halogen atoms' may include fluorine, chlorine, bromine and iodine atoms. An "alkyl group" includes both straight, pulverized and cyclic carbon chains having 1 to 8 carbon atoms. Specifically, the alkyl group may include a methyl group, ethyl group, propyl group, isopropyl group, butyl group, isobutyl group, t -butyl group, cyclopentyl group, cyclohexyl group and the like. "Alkoxy group" means an alkyl group of carbon connected to oxygen, wherein alkyl is as defined above. "Benzyl group" refers to a functional group capable of forming a covalent bond to another atom by methylene substituted by a phenyl group and substituted methylene carbon.

In the compound represented by Chemical Formula 1 according to the present invention, preferably, R ¹ and R ² are the same as or different from each other, and are a phenyl group, 3-chlorophenyl group, 4-chlorophenyl group, 2,3-dichlorophenyl group, 2 , 3-difluorophenyl group, 4- (tripulouromethyl) phenyl group, 2-methylphenyl group, 3-methylphenyl group, 4-methylphenyl group, 4- t -butylphenyl group, 4-cyclohexylphenyl group, 2,3 -Dimethylphenyl group, 2,4-dimethylphenyl group, 2,4,5-trimethylphenyl group, 4-methoxyphenyl group, benzyl group, 4- t -butylbenzyl group, 2,4,6-trimethylbenzyl group, 2 It is a compound which shows a 3, 4- trimethoxy benzyl group, a (benzo [ d ] [1, 3] diosol-5-yl) methyl group, a furyl group, a furan-2-ylmethyl group, or a pyridin-2-yl group. .

In addition, the present invention is a 1,6-disubstituted-3-amino-4,5,6,7-tetrahydro-1 H -pyrazolo [3,4- c ] pyridin-7-one compound represented by the formula (1) Or a pharmaceutical composition for treating and preventing cancer containing the pharmaceutically acceptable salt thereof as an active ingredient.

In addition, the present invention is a 1,6-disubstituted-3-amino-4,5,6,7-tetrahydro-1 H -pyrazolo [3,4- c ] pyridin-7-one compound represented by the formula (1) Or an anticancer agent containing a pharmaceutically acceptable salt thereof as an active ingredient.

The compound represented by the formula (1) according to the present invention is specifically exemplified as follows:

3-amino-1-[{4- (2,3-dimethylphenyl) piperazin-1-yl} ethanoyl] -6- N- ( p- tolyl) -4,5,6,7-tetrahydro- 1 H- pyrazolo [3,4- c ] pyridin-7-one (Compound No. 1),

3-amino-1-[{4- (2,3-dimethylphenyl) piperazin-1-yl} propanoyl] -6- N- ( p- tolyl) -4,5,6,7-tetrahydro -1 H- pyrazolo [3,4- c ] pyridin-7-one (Compound No. 2),

3-amino-1-{(4-phenylpiperazin-1-yl) propanoyl} -6- N- ( p- tolyl) -4,5,6,7-tetrahydro-1 H- pyrazolo [ 3,4- c ] pyridin-7-one (Compound No. 3),

3-amino-1-[{4- (2,4-dimethylphenyl) piperazin-1-yl} propanoyl] -6- N- ( p- tolyl) -4,5,6,7-tetrahydro -1 H- pyrazolo [3,4- c ] pyridin-7-one (Compound No. 4),

3-amino-1-[{4-((benzo [ d ] [1,3] dioxol-5-yl) methyl) piperazin-1-yl} propanoyl] -6- N- ( p- tolyl ) -4,5,6,7-tetrahydro-1 H- pyrazolo [3,4- c ] pyridin-7-one (Compound No. 5),

3-amino-1-[{4- (2,4,6-trimethylbenzyl) piperazin-1-yl} propanoyl] -6- N- ( p- tolyl) -4,5,6,7- Tetrahydro-1 H- pyrazolo [3,4- c ] pyridin-7-one (Compound No. 6),

3-amino-1-[{4- (4- t- butylbenzyl) piperazin-1-yl} propanoyl] -6- N- ( p- tolyl) -4,5,6,7-tetrahydro -1 H- pyrazolo [3,4- c ] pyridin-7-one (Compound No. 7),

3-amino-1-[{4- (2,3,4-trimethoxybenzyl) piperazin-1-yl} propanoyl] -6- N- ( p- tolyl) -4,5,6, 7-tetrahydro-1 H- pyrazolo [3,4- c ] pyridin-7-one (Compound No. 8),

3-amino-1-[{4- (4-chlorophenyl) piperazin-1-yl} propanoyl] -6- N- ( p- tolyl) -4,5,6,7-tetrahydro-1 H- pyrazolo [3,4- c ] pyridin-7-one (Compound No. 9),

3-amino-1-[{4- (4-methoxyphenyl) piperazin-1-yl} propanoyl] -6- N- ( p- tolyl) -4,5,6,7-tetrahydro- 1 H- pyrazolo [3,4- c ] pyridin-7-one (Compound No. 10),

3-amino -1 - {(N-benzyl) piperazin-1-yl} -6- N - (p- methoxyphenyl) 4,5,6,7-tetrahydro-pyrazolo [3,4 - c ] pyridin-7-one (Compound No. 11),

3-amino-1-[{4- (2,4-dimethylphenyl) piperazin-1-yl} propanoyl] -6- N- ( p -methoxyphenyl) -4,5,6,7- Tetrahydro-1 H- pyrazolo [3,4- c ] pyridin-7-one (Compound No. 12),

3-amino-1-[{4- (2,4,6-trimethylbenzyl) piperazin-1-yl} propanoyl] -6- N- ( p -methoxyphenyl) -4,5,6, 7-tetrahydro-1 H- pyrazolo [3,4- c ] pyridin-7-one (Compound No. 13),

3-amino-1-[{4- (2,3,4-trimethoxybenzyl) piperazin-1-yl} propanoyl] -6- N- ( p -methoxyphenyl) -4,5, 6,7-tetrahydro-1 H- pyrazolo [3,4- c ] pyridin-7-one (Compound No. 14),

3-amino-1-[{4- (2,4-dimethylphenyl) piperazin-1-yl} propanoyl] -6- N -phenyl-4,5,6,7-tetrahydro-1 H- Pyrazolo [3,4- c ] pyridin-7-one (Compound No. 15),

3-amino-1-[{4- (2,4,6-trimethylbenzyl) piperazin-1-yl} propanoyl] -6- N -phenyl-4,5,6,7-tetrahydro-1 H- pyrazolo [3,4- c ] pyridin-7-one (Compound No. 16),

3-amino-1-[{4- (2,4-dimethylphenyl) piperazin-1-yl} propanoyl] -6- N -benzyl-4,5,6,7-tetrahydro-1 H- Pyrazolo [3,4- c ] pyridin-7-one (Compound No. 17),

3-amino-1-{(4-phenylpiperazin-1-yl) propanoyl} -6- N- { p- (dimethylamino) phenyl} -4,5,6,7-tetrahydro-1 H- pyrazolo [3,4- c ] pyridin-7-one (Compound No. 18),

3-amino-1-[{4- (4-methoxyphenyl) piperazin-1-yl} propanoyl] -6- N- { p- (dimethylamino) phenyl} -4,5,6, 7-tetrahydro-1 H- pyrazolo [3,4- c ] pyridin-7-one (Compound No. 19),

3-amino-1-{(4-phenylpiperazin-1-yl) propanoyl} -6- N -{(furan-2-yl) methyl} -4,5,6,7-tetrahydro-1 H- pyrazolo [3,4- c ] pyridin-7-one (Compound No. 20),

3-amino-1-[{4- (4-methoxyphenyl) piperazin-1-yl} propanoyl] -6- N -{(furan-2-yl) methyl} -4,5,6, 7-tetrahydro-1 H- pyrazolo [3,4- c ] pyridin-7-one (Compound No. 21),

3-amino-1- {4- (2-pyridinylpiperazin-1-yl) propanoyl} -6- Np- tolyl-4,5,6,7-tetrahydro-1 H -pyrazolo [3 , 4- c ] pyridin-7-one (Compound No. 22),

3-amino-1- {4- (2-pyridinylpiperazin-1-yl) propanoyl} -6- N -{(furan-2-yl) methyl} -4,5,6,7-tetra Hydro-1 H -pyrazolo [3,4- c ] pyridin-7-one (Compound No. 23),

3-amino-1-[{4- (4-trifluorofluoromethylphenyl) piperazin-1-yl} propanoyl] -6- N- ( p- tolyl) -4,5,6,7-tetra Hydro-1 H -pyrazolo [3,4- c ] pyridin-7-one (Compound No. 24),

3-amino-1-[{4- (2,4-dimethylphenyl) piperazin-1-yl} propanoyl] -6- N -{(furan-2-yl) methyl} -4,5,6 , 7-tetrahydro-1 H -pyrazolo [3,4- c ] pyridin-7-one (Compound No. 25),

3-amino-1-[{4- (2,4,5-trimethylphenyl) piperazin-1-yl} propanoyl] -6- N- ( p- tolyl) -4,5,6,7- Tetrahydro-1 H -pyrazolo [3,4- c ] pyridin-7-one (Compound No. 26),

3-amino-1-[{4- (4- t- butylphenyl) piperazin-1-yl} propanoyl] -6- N- ( p- tolyl) -4,5,6,7-tetrahydro -1 H -pyrazolo [3,4- c ] pyridin-7-one (Compound No. 27),

3-amino-1-[{4- (cyclohexylphenyl) piperazin-1-yl} propanoyl] -6- N- ( p- tolyl) -4,5,6,7-tetrahydro-1 H -Pyrazolo [3,4- c ] pyridin-7-one (Compound No. 28).

In addition, the present invention is a 1,6-disubstituted-3-amino-4,5,6,7-tetrahydro-1 H -pyrazolo [3,4- c ] pyridin-7-one compound represented by the formula (1) It includes the manufacturing method of.

As shown in Scheme 1, the preparation method according to the present invention is an alkylation reaction of the 3-amino compound represented by the following formula (2) and the haloalkylcarbonyl peparagine compound represented by the following formula (3), A 1,6-disubstituted-3-amino-4,5,6,7-tetrahydro-1 H -pyrazolo [3,4- c ] pyridin-7-one compound can be prepared.

[Reaction Scheme 1]

In Scheme 1, n, R ¹ , and R ² are each as defined in Chemical Formula 1, and X represents a halogen atom.

The alkylation reaction can be carried out by refluxing in the presence of a base. In this case, the base used may be a conventional organic base selected from aliphatic alkylamines including triethylamine, aromatic amines including pyridine, or the like, or inorganic bases selected from hydroxides, carbonates, hydrogencarbonates, sulfates and the like of alkali or alkaline earth metals. Also good. As the reaction solvent, an inert organic solvent which does not affect the reaction may be used as an organic solvent commonly used in the art. As an organic solvent, For example, ethers, such as diethyl ether; Lower alcohols having 1 to 6 carbon atoms such as methanol, ethanol and propanol; Tetrahydrofuran; Halogenated compounds such as chloroform, methylene chloride and the like; Nitrile compounds, such as acetonitrile, etc. can be used, or it is also possible to use these mixed solvents selected from these. The reaction temperature is preferably carried out by heating at a temperature in the range of 30 ° C. to 80 ° C. as the reflux temperature of the solvent used.

In addition, the present invention, the 3-amino compound represented by the formula (2) used in the preparation method according to Scheme 1 is a novel intermediate compound, which includes it as a right scope.

The 3-amino compound represented by Chemical Formula 2 may be prepared by performing a manufacturing process as shown in Scheme 2 below.

Scheme 2

In Scheme 2, R ¹ is as defined in Formula 1.

According to Scheme 2, the method for preparing 3-amino compound represented by Formula 2 is:

Iii) refluxing the amine compound represented by Chemical Formula 4 with 4-bromobutyronitrile compound represented by Chemical Formula 5 under basic conditions to prepare 4-aminobutanenitrile compound represented by Chemical Formula 6;

Ii) a 4-aminobutanenitrile compound represented by the formula (6) by a condensation reaction with a β-diketone compound represented by the formula (7) by a condensation reaction under a basic condition, and the piperidine compound represented by the formula (8) Manufacturing process; And

Iii) cyclizing the piperidine compound represented by Chemical Formula 8 with hydrazine under acetic acid and an alcohol mixed solvent to prepare a 3-amino compound represented by Chemical Formula 2; It is made, including.

The amine compound represented by Formula 4 used as a starting material in the preparation method of Scheme 2 may be purchased by commercially available or prepared by a method known in the literature. The base used in Scheme 2 may be selected from conventional inorganic bases or organic bases used in the art as mentioned above. As the solvent, an inert organic solvent which does not affect the reaction may be used as an organic solvent commonly used in the art. Specific examples of the organic solvent that can be used in the present invention include ethers such as diethyl ether; Lower alcohols having 1 to 6 carbon atoms such as methanol, ethanol and propanol; Tetrahydrofuran; Halogenated compounds such as chloroform, methylene chloride and the like; Nitrile compounds, such as acetonitrile, etc. can be used. The reaction temperature can be carried out in the reflux temperature range of -30 ℃ to the solvent used, more specifically in the temperature range of room temperature to 120 ℃, even more specifically can be carried out within 30 to 80 ℃.

In addition, the haloalkylcarbonyl peparagine compound represented by Chemical Formula 3 used as a starting material in the preparation method according to Scheme 1 may be prepared by performing the preparation method as shown in Scheme 3 below.

Scheme 3

In Scheme 3, n, R ¹ , and R ² are each as defined in Chemical Formula 1, and X represents a halogen atom.

In the preparation method according to Scheme 3, the piperazine compound represented by the formula (9) and the haloacyl halide compound represented by the following formula (10) are amidated to prepare a haloalkylcarbonyl peparazine compound represented by the formula (3). do. The amidation reaction can be carried out in the presence of an N, N -dimethylaminopyridine (DMAP) catalyst and an amine base. The amine base may use a conventional organic base selected from aliphatic alkylamines including triethylamine, aromatic amines including pyridine and the like. As a reaction solvent, the organic solvent used normally, For example, C1-C6 lower alcohols, such as diethyl ether, methanol, ethanol, a propanol; Tetrahydrofuran; Halogenated compounds such as chloroform, methylene chloride and the like; Nitrile compounds, such as acetonitrile, etc. can be used, or it is also possible to use these mixed solvents selected from these. The reaction temperature is preferably performed in a temperature range of -20 ° C to room temperature, preferably in a temperature range of 0 ° C to 20 ° C.

In addition, the compound represented by Formula 1 according to the present invention has a growth inhibitory effect on various cancer cell lines, and thus may be used as an active ingredient of an anticancer agent. Therefore, the present invention includes the anticancer agent or the pharmaceutical composition for anticancer agent containing the compound represented by the formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient as a scope. In order to measure the anticancer activity of the compounds according to the present invention, the degree of activity inhibition for each anticancer cell line was measured.

Pharmaceutically acceptable salts of the compounds represented by the formula (1) used in the preparation of the pharmaceutical composition according to the present invention can be prepared by methods commonly known in the art. The invention places no particular limitation on its selection. Pharmaceutically acceptable salts include, for example, salts with nontoxic inorganic acids such as hydrochloric acid, bromic acid, sulfuric acid, sodium hydrogen sulfate, phosphoric acid, nitric acid, carbonic acid; Formic acid, acetic acid, propionic acid, oxalic acid, succinic acid, glycolic acid, stearic acid, lactic acid, maleic acid, malonic acid, tartaric acid, citric acid, benzoic acid, paratoluenesulfonic acid, methanesulfonic acid, gluconic acid, lactic acid, gesty acid, fumaric acid, lactoic acid Salts with non-toxic organic acids such as ionic acid, salicylic acid, or acetylsalicylic acid (aspirin); Salts with amino acids such as glycine, alanine, vanillin, isoleucine, serine, cysteine, cystine, aspartic acid, glutamine, lysine, arginine, tyrosine, proline and the like; Salts with sulfonic acids such as methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, toluenesulfonic acid and the like; Metal salts by reaction with alkali metals such as sodium and potassium; Or salts with ammonium ions.

In addition, the pharmaceutical composition of the present invention is a conventional formulation in the pharmaceutical field by adding a conventional non-toxic pharmaceutically acceptable carrier, adjuvant and excipient to the compound represented by the formula (1) or pharmaceutically acceptable salts thereof For example, it may be prepared by oral or parenteral administration such as tablets, capsules, troches, solutions, suspensions, and the like, and may be used for the prevention and treatment of various types of tumors.

Excipients that can be used in the pharmaceutical compositions of the present invention may include sweeteners, binders, solubilizers, dissolution aids, wetting agents, emulsifiers, isotonic agents, adsorbents, disintegrants, antioxidants, preservatives, lubricants, fillers, fragrances and the like. For example, lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, glycine, silica, talc, stearic acid, sterin, magnesium stearate, magnesium aluminum silicate, starch, gelatin, tragacanth rubber, arginine acid, sodium Alginate, methyl cellulose, sodium carboxymethyl cellulose, agar, water, ethanol, polyethylene glycol, polyvinylpyrrolidone, sodium chloride, calcium chloride, orange essence, strawberry essence, vanilla flavor and the like.

In addition, the dosage of the compound represented by Formula 1 according to the present invention to the human body may vary depending on the age, weight, sex, dosage form, health condition and degree of disease of the patient, based on an adult patient weighing 70 kg In general, 0.01 mg to 5000 mg per day, and may be dividedly administered once to several times a day at regular intervals according to the judgment of a doctor or pharmacist.

Hereinafter, the present invention will be described in detail with respect to the preparation method and efficacy of the compound according to the present invention through Examples and Experimental Examples. However, these Examples and Experimental Examples are only for helping the understanding of the present invention, and the scope of the present invention is not limited thereto.

[Example]

Reference Example 1. 4- ( p- tolylamino) butanenitrile

After dissolving p- toluidine (10.407 g, 97.116 mmol) in anhydrous acetonitrile, potassium carbonate (49.9 g, 361.272 mmol) was added thereto, followed by stirring at room temperature for 10 minutes. 4-Bromobutyronitrile (10.1 mL, 101.972 mmol) was slowly added to the reaction mixture, followed by reaction at 100 ° C. for 2 days. After the mixture was cooled to room temperature, the produced solid was removed by filtration, and the filtrate was concentrated under reduced pressure to obtain 13.23 g (78.2%) of the title compound. It was used for the next reaction without further purification.

¹ H NMR (CDCl ₃ , 300 MHz) δ 7.04 (d, J = 8.0 Hz, 2H), 6.57 (d, J = 8.3 Hz, 2H), 3.62 (s, 1H, NH), 3.29 (q, J = 6.3 Hz, 2H), 2.47 (t, J = 7.0 Hz, 2H), 2.28 (s, 3H), 1.96 (t, J = 6.8 Hz, 2H)

Reference Example 2. 4- (4-methoxyphenylamino) butanenitrile

p -anisidine (4.6826 g, 38.020 mmol), potassium carbonate (19.5 g, 141.434 mmol) and 4-bromobutyronitrile (3.95 mL, 39.921 mmol) were reacted in the same manner as in Reference Example 1 above. 5.6 g (29.436 mmol, 77.5%) of the title compound were obtained.

¹ H NMR (CDCl ₃ , 300 MHz) δ 6.81 (d, J = 8.9 Hz, 2H), 6.61 (d, J = 8.9 Hz, 2H), 3.77 (s, 3H), 3.42 (s, 1H, NH) , 3.28 (t, J = 6.6 Hz, 2H), 2.49 (t, J = 7.1 Hz, 2H), 2.02-1.93 (m, 2H)

Reference Example 3. 4- (benzylamino) butanenitrile

Benzylamine (5.02 mL, 46 mmol) and calcium carbonate (23.65 g, 171.12 mmol) dissolved in anhydrous acetonitrile (115 mL) were stirred at room temperature for 10 minutes, and 4-bromononitrile (4.78 mL, 48.3 mmol) was added thereto. After the reaction was carried out at 100 ° C. for 2 days. The reaction mixture was separated in the same manner as in Reference Example 1 to obtain 6.69 g (38.395 mmol; 83.5%) of the target compound in a liquid state.

¹ H NMR (CDCl ₃ , 300 MHz) δ 7.37-7.29 (m, 5H), 3.79 (s, 2H), 2.77 (t, J = 6.6 Hz, 2H), 2.48 (t, J = 7.1 Hz, 2H) , 1.88-1.79 (m, 2H)

Reference Example 4. 2,3-Dioxo-1- p- tolylpiperidine-4-carbonitrile

4- ( p- tolylamino) butanenitrile (13.23 g, 75.929 mmol) was dissolved in 189.8 mL of ethanol and diethyl oxalic acid (10.3 mL, 75.929 mmol) was added, followed by sodium ethoxide (34.0 mL, 91.115). mmol) was added and refluxed for 2 days. The reaction solution was cooled to room temperature, and the resulting solid was filtered to give 8.94 g (39.167 mmol, 51.6%) of the title compound as a yellow solid.

¹ H NMR (CDCl ₃ , 300 MHz) δ 7.65 (s, 1H), 7.25 (d, J = 8.5 Hz, 2H), 7.18 (d, J = 8.3 Hz, 2H), 3.92 (t, J = 6.9 Hz , 2H), 2.76 (t, J = 6.9 Hz, 2H), 2.38 (s, 3H)

Reference Example 5. 1- (4-methoxyphenyl) -2,3-dioxopiperidine-4-carbonitrile

Using 4- (4-methoxyphenylamino) butanenitrile (5.6 g, 29.436 mmol), diethyl oxalic acid (4.19 mL, 30.908 mmol) and sodium ethoxide (14.3 mL, 38.267), the same as in Reference Example 4 above Reaction was carried out to give 4.87 g (19.938 mmol, 67.7%) of the title compound as a yellow solid.

¹ H NMR (DMSO, 300 MHz) δ 7.26 (d, J = 8.7 Hz, 2H), 6.96 (d, J = 8.7 Hz, 2H), 3.79 (t, J = 6.8 Hz, 2H), 3.76 (s, 3H), 2.61 (t, J = 6.7 Hz, 2H).

Reference Example 6. 1-benzyl-2,3-dioxopiperidine-4-carbonitrile

Reaction in the same manner as in Reference Example 4, using 4- (benzylamino) butanenitrile (6.69 g, 38.395 mmol), diethyl oxalic acid (5.46 mL, 40.314 mmol) and sodium ethoxide (18.6 mL, 49.913 mmol) This yielded 4.1 g (17.962 mmol, 46.8%) of the title compound as a yellow solid.

¹ H NMR (CDCl ₃ , 300 MHz) δ 7.84 (s, 1H), 7.41-7.31 (m, 3H), 7.30-7.25 (m, 2H), 4.65 (s, 2H), 3.42 (t, J = 7.1 Hz, 2H), 2.56 (t, J = 7.1 Hz, 2H)

Reference Example 7. 3-Amino-5, 6 - dihydro-6- p- tolyl-1 H- pyrazolo [3,4- c ] pyridin-7 ( 4H ) -one

2,3-dioxo-1- p- tolylpiperidine-4-carbonitrile (3.56 g, 15.597 mmol) was added to anhydrous ethanol (52.0 mL), dissolved by heating to 60 ° C, and hydrazine hydrochloride (1.06 mL) at room temperature. , 21.836 mmol) and glacial acetic acid (1.34 mL, 23.396 mmol) were added and allowed to react for 2 days. After distilling off ethanol, the resulting solid was filtered and washed two more times with NaHCO ₃ to completely remove acetic acid to obtain 3.25 g (13.414 mmol, 86.0%) of the title compound as a white solid.

¹ H NMR (DMSO, 300 MHz) δ 12.24 (s, 1H, NH), 7.23-7.16 (m, 4H), 4.84 (s, 2H, NH ₂ ), 3.87 (t, J = 6.6 Hz, 2H), 2.69 (t, J = 6.6 Hz, 2H), 2.30 (s, 3H)

Reference Example 8. 3-Amino-5, 6-dihydro-6- (4-methoxyphenyl) -1 H- pyrazolo [3,4- c ] pyridin-7 ( 4H ) -one

1- (4-methoxyphenyl) -2,3-dioxopiperidine-4-carbonitrile (1.63 g, 6.673 mmol), hydrazine hydrate (0.45 mL, 9.342 mmol) and glacial acetic acid (0.57 mL, 10.009 mmol) In the same manner as in Reference Example 7 to obtain 1.57 g (6.079 mmol, 91.3%) of the title compound as a white solid.

¹ H NMR (DMSO, 300 MHz) δ 12.26 (s, 1H, NH), 7.24 (d, J = 8.7 Hz, 2H), 6.94 (d, J = 8.8 Hz, 2H), 4.81 (s, 2H, NH ₂ ), 3.85 (t, J = 6.5 Hz, 2H), 3.76 (s, 3H), 2.69 (t, J = 6.6 Hz, 2H)

Reference Example 9. 3-Amino-6-benzyl-5, 6-dihydro-1 H- pyrazolo [3,4- c ] pyridin-7 ( 4H ) -one

Reference Example Using 1-benzyl-2,3-dioxopiperidine-4-carbonitrile (4.1 g, 17.962 mmol), hydrazine hydrochloride (1.22 mL, 25.146 mmol) and glacial acetic acid (1.54 mL, 26.943 mmol) 7.23 g (17.459 mmol, 97.2%) of the title compound was obtained as a white solid.

¹ H NMR (CDCl ₃ , 300 MHz) δ 7.38-7.27 (m, 5H), 4.74 (s, 2H), 4.69 (s, 2H, NH ₂ ), 3.52 (t, J = 6.8 Hz, 2H), 2.64 (t, J = 6.8 Hz, 2H)

Reference Example 10 1- (4 -t- butylphenyl) piperazine

1-bromo-4- t- butyl benzene (2.38 mL, 14 mmol), piperazine (1.8 g, 21 mmol), sodium t- butoxide (1.95 g, 20.3 mmol) and dichlorobis (tri- o- Tolylphosphine) palladium (II) (330.1 mg, 0.42 mmol) was dissolved in anhydrous toluene and heated at 100 ° C. for 3 hours. After confirming complete disappearance of 1-bromo-4 -t- butyl benzene by TLC, the reaction solution was filtered through Celite. The filtrate was concentrated and eluted by column chromatography (CH ₂ Cl ₂ / MeOH, 1: 1) to give 1.47 g (6.732 mmol, 48.2%) of the title compound.

¹ H NMR (CDCl ₃ , 300 MHz) δ 7.31 (d, J = 8.6 Hz, 2H), 6.90 (d, J = 8.6 Hz, 2H), 3.14 (br, 4H), 3.04 (br, 4H), 1.57 (s, 1H, NH), 1.31 (s, 9H)

¹³ C NMR (CDCl ₃ , 75 MHz) δ 149.5, 142.4, 125.8, 115.7, 50.6, 46.3, 33.9, 31.4

Reference Example 11. 1- (2,4,5-trimethylphenyl) piperazine

5-Bromo-1,2,4-trimethylbenzene (2.5 g, 12.556 mmol), piperazine (4.32 g, 50.224 mmol), sodium t- butoxide (1.75 g, 18.206 mmol) in the same manner as in Reference Example 10 above. ) And dichlorobis (tri- o- tolylphosphine) palladium (II) (295.5 mg, 0.37 mmol) were reacted and separated and purified by column chromatography (CH ₂ Cl ₂ / MeOH, 1: 1). This resulted in 1.03 g (5.041 mmol, 40.2%) of the title compound.

¹ H NMR (CDCl ₃ , 300 MHz) δ 6.97 (s, 1H), 6.83 (s, 1H), 3.14 (br, 4H), 2.96 (br, 4H), 2.24 (s, 3H), 2.22 (s, 3H), 2.19 (s, 3H)

Reference Example 12 1- (4-cyclohexylphenyl) piperazine

In the same manner as in Reference Example 10, 1-bromo-4-cyclohexyl benzene (3.34 mL, 18 mmol), piperazine (2.01 g, 23.4 mmol), sodium t- butoxide (2.51 g, 26.1 mmol) and di Chlorobis (tri- o- tolylphosphine) palladium (II) (424.4 mg, 0.54 mmol) was reacted, eluted with column chromatography (CH ₂ Cl ₂ / MeOH, 1: 1), and purified by purification. 2.01 g (8.224 mmol, 45.7%) was obtained.

¹ H NMR (CDCl ₃ , 300 MHz) δ 7.13 (d, J = 8.5 Hz, 2H), 6.88 (d, J = 8.6 Hz, 2H), 3.13-3.10 (m, 4H), 3.05-3.02 (m, 4H), 2.44 (br, 1H), 1.85 (br, 4H), 1.74 (d, J = 12.1 Hz, 1H), 1.46-1.21 (m, 5H)

Reference Example 13. 3-Chloro-1- (4- (4-methoxyphenyl) piperazin-1-yl) propan-1-one

 1- (4-methoxyphenyl) piperazine dihydrochloride (1.85 g, 6.976 mmol) was dissolved in methylene chloride, and then triethylamine (1.9 mL, 13.952 mmol) was added dropwise at 0 ° C., and stirred for 30 minutes. 3-chloropropionyl chloride (0.67 mL, 6.976 mmol) was added to the reaction solution, and the mixture was stirred at room temperature for 1 hour. The reaction solution was added with methanol and extracted with methylene chloride. The organic layer was dried over a desiccant, filtered and the filtrate was concentrated and eluted by column chromatography (EtOAC / Hexane, 4: 1) to obtain 1.76 g (6.224 mmol, 89.3%) of the title compound as a white solid.

¹ H NMR (CDCl ₃ , 300 MHz) δ 6.93-6.85 (m, 4H), 3.87 (t, J = 7.0 Hz, 2H), 3.80 (br, 2H), 3.79 (s, 3H), 3.64 (br, 2H), 3.06 (br, 4H), 2.86 (t, J = 7.0 Hz, 2H)

Reference Example 14. 3-Chloro-1- (4- (4-chlorophenyl) piperazin-1-yl) propan-1-one

1- (4-chlorophenyl) piperazine dihydrochloride (1.88 g, 6.973 mmol) was dissolved in methylene chloride, then triethylamine (1.9 mL, 13.946 mmol) was slowly added at 0 ° C. and stirred for 5 minutes. 3-chloropropionyl chloride (0.67 mL, 6.973 mmol) was added to the reaction solution, and stirred at room temperature. After extraction with methylene chloride, the organic layer was dried with a desiccant, filtered and the filtrate was concentrated and eluted by column chromatography (EtOAC / Hexane, 4: 1) to purify. The title compound in white solid 1.03 g (3.586 mmol, 51.5%). )

¹ H NMR (CDCl ₃ , 300 MHz) δ 7.24 (d, J = 9.0 Hz, 2H), 6.85 (d, J = 8.9 Hz, 2H), 3.87 (t, J = 6.9 Hz, 2H), 3.81 (br , 2H), 3.65 (br, 2H), 3.16 (br, 4H), 2.86 (t, J = 6.9 Hz, 2H)

Reference Example 15 3-chloro-1- (4-phenylpiperazin-1-yl) propan-1-one

In the same manner as in Reference Example 13, using 1-phenylpiperazine (1.82 mL, 12 mmol) and 3-chloropropionyl chloride (0.67 mL, 6.976 mmol), 2.41 g (9.535 mmol, 80.35%).

¹ H NMR (CDCl ₃ , 300 MHz) δ 7.30 (t, J = 8.1 Hz, 2H), 6.96-6.90 (m, 3H), 3.87 (t, J = 7.0 Hz, 2H), 3.81 (br, 2H) , 3.65 (br, 2H), 3.19 (br, 4H), 2.87 (t, J = 7.0 Hz, 2H)

Reference Example 16 3-Chloro-1- (4- (2,4-dimethylphenyl) piperazin-1-yl) propan-1-one

Using the 1- (2,4-dimethylphenyl) piperazine (2 g, 10.510 mmol) and 3-chloropropionyl chloride (1.0 mL, 10.510 mmol) in the same manner as in Reference Example 13, the purpose of the white solid state 1.72 g (6.125 mmol, 58.3%) were obtained.

¹ H NMR (CDCl ₃ , 300 MHz) δ 7.04 (s, 1H), 6.99 (d, J = 8.0 Hz, 1H), 6.89 (d, J = 8.0 Hz, 1H), 3.87 (t, J = 7.0 Hz , 2H), 3.78 (t, J = 4.7 Hz, 2H), 3.62 (t, J = 4.9 Hz, 2H), 2.91-2.85 (m, 6H), 2.30 (s, 3H), 2.29 (s, 3H)

Reference Example 17 3-Chloro-1- (4- (pyridin-2-yl) piperazin-1-yl) propan-1-one

In the same manner as in Reference Example 13, 1- (pyridin-2-yl) piperazine (1.8 mL, 11.822 mmol) and 3-chloropropionyl chloride (1.13 mL, 11.822 mmol) were used to obtain the target compound as a white solid. 1.3 g (5.123 mmol, 43.3%) was obtained.

¹ H NMR (CDCl ₃ , 300 MHz) δ 8.21 (d, J = 6.6 Hz, 1H), 7.52 (t, J = 7.8 Hz, 1H), 6.71-6.65 (m, 2H), 3.87 (t, J = 6.9 Hz, 2H), 3.78 (t, J = 5.3 Hz, 2H), 3.66-3.59 (m, 4H), 3.52 (t, J = 5.3 Hz, 2H), 2.87 (t, J = 6.9 Hz, 2H)

Reference Example 18 3-Chloro-1- (4- (4- (trifuluromethyl) phenyl) piperazin-1-yl) propan-1-one

In the same manner as in Reference Example 13, N- (α, α, α - tripulouro- p- tolyl) piperazine (500 mg, 2.171 mmol) and 3-chloropropionyl chloride (208.8 mg, 2.171 mmol) were added. To give 492.9 mg (1.536 mmol, 70.7%) of the title compound as a white solid.

¹ H NMR (CDCl ₃ , 300 MHz) δ 7.52 (d, J = 8.7 Hz, 2H), 6.94 (d, J = 8.6 Hz, 2H), 3.87 (t, J = 6.9 Hz, 2H), 3.82 (t , J = 5.2 Hz, 2H), 3.67 (t, J = 5.1 Hz, 2H), 3.34-3.27 (m, 4H), 2.87 (t, J = 6.9 Hz, 2H)

Reference Example 19 3-chloro-1- (4- (2,4,5-trimethylphenyl) piperazin-1-yl) propan-1-one

In the same manner as in Reference Example 13, 1- (2,4,5-trimethylphenyl) piperazine (1.03 g, 5.041 mmol) and 3-chloropropionyl chloride (484.9 mg, 5.041 mmol) were reacted and column chromatography Elution with (EtOAC / Hexane, 1: 2) separated and purified to give 925 mg (3.137 mmol, 62.5%) of the title compound in the liquid state.

¹ H NMR (CDCl ₃ , 300 MHz) δ 6.98 (s, 1H), 6.77 (s, 1H), 3.87 (t, J = 7.0 Hz, 2H), 3.78 (t, J = 4.8 Hz, 2H), 3.61 (t, J = 4.9 Hz, 2H), 2.91-2.84 (m, 6H), 2.27 (s, 3H), 2.22 (s, 3H), 2.20 (s, 3H)

Reference Example 20 1- (4- (4 -t- butylphenyl) piperazin-1-yl) -3-chloropropan-1-one

In the same manner as in Reference Example 13, 1- (4 -t- butylphenyl) piperazine (1.47 g, 6.732 mmol) and 3-chloropropionyl chloride (647.5 mg, 6.732 mmol) were reacted and column chromatography (EtOAC) was performed. Eluting with: Hexane: CH ₂ Cl ₂ , 1: 1: 5) to obtain 1.59 g (5.148 mmol, 76.4%) of the title compound in the liquid state.

¹ H NMR (CDCl ₃ , 300 MHz) δ 7.32 (d, J = 8.8 Hz, 2H), 6.89 (d, J = 8.8 Hz, 2H), 3.87 (t, J = 7.0 Hz, 2H), 3.80 (t , J = 5.1 Hz, 2H), 3.64 (t, J = 5.1 Hz, 2H), 3.19-3.13 (m, 4H), 2.87 (t, J = 7.0 Hz, 2H), 1.31 (s, 9H)

Reference Example 21 3-chloro-1- (4- (4-cyclohexylphenyl) piperazin-1-yl) propan-1-one

In the same manner as in Reference Example 13, 1- (4-cyclohexylphenyl) piperazine (2.01 g, 8.224 mmol) and 3-chloropropionyl chloride (0.79 mL, 8.224 mmol) were reacted, and column chromatography (EtOAC: Hexane: CH ₂ Cl ₂ , 1: 1: 5) eluted and purified to obtain 2.2 g (6.569 mmol) of the target compound in the form of a white solid.

¹ H NMR (CDCl ₃ , 300 MHz) δ 7.14 (d, J = 8.6 Hz, 2H), 6.88 (d, J = 8.6 Hz, 2H), 3.86 (t, J = 7.0 Hz, 2H), 3.80 (t , J = 5.1 Hz, 2H), 3.64 (t, J = 5.1 Hz, 2H), 3.18-3.12 (m, 4H), 2.86 (t, J = 7.0 Hz, 2H), 2.45 (br, 1H), 1.84 (br, 4H), 1.75 (d, J = 12.0 Hz, 1H), 1.46-1.22 (m, 5H)

Reference Example 22 1- N - t -Butoxycarbonyl-3-amino-4,5,6,7-tetrahydro-6- N- ( p- tolyl) pyrazolo [3,4- c ] pyridine- 7-on

Synthesized from the Reference Example 7 3-Amino-5,6-dihydro -6- p- tolyl -1 H- pyrazolo [3,4 -c] pyridin -7 (4 H) - one (1 g, 4.127 mmol), and potassium carbonate (1.1 g, 8.254 mmol) in anhydrous tetrahydrofuran (10.3 mL) was dissolved di t - Insert the butyl dicarbonate (990.6 mg, 4.539 mmol) was reacted for 2 days at room temperature. The reaction mixture was extracted with methylene chloride, the organic layer was dried over MgSO _4, and concentrated under reduced pressure. Separation and purification gave 987.8 mg (2.885 mmol, 70.1%) of the title compound.

¹ H NMR (CDCl ₃ , 300 MHz) δ 7.20 (s, 4H), 5.31 (s, 2H, NH ₂ ), 3.93 (t, J = 6.4 Hz, 2H), 2.73 (t, J = 6.4 Hz, 2H ), 2.35 (s, 3H), 1.65 (s, 9H)

Reference Example 23 1- N - t -Butoxycarbonyl-3-amino-4,5,6,7-tetrahydro-6- N- (4-methoxyphenyl) pyrazolo [3,4- c ] Pyridin-7-one

3-amino-5,6-dihydro-6- (4-methoxyphenyl) -1 H- pyrazolo [3,4- c ] pyridine- synthesized from Reference Example 8 in the same manner as in Reference Example 22 above The target compound was obtained using 7 ( 4H ) -one.

¹ H NMR (CDCl ₃ , 300 MHz) δ 7.24 (d, J = 8.9 Hz, 2H), 6.93 (d, J = 8.9 Hz, 2H), 5.26 (s, 2H, NH ₂ ), 3.93 (t, J = 6.4 Hz, 2H), 3.82 (s, 3H), 2.74 (t, J = 6.4 Hz, 2H), 1.66 (s, 9H)

Example 1. 3-Amino-1-[{4- (2,3-dimethylphenyl) piperazin-1-yl} ethanoyl] -6- N- ( p- tolyl) -4,5,6,7 -1 H- pyrazolo [3,4- c ] pyridin-7-one (Compound No. 1)

1- N - t -butoxycarbonyl-3-amino-4,5,6,7-tetrahydro-6- N- ( p- tolyl) pyrazolo [3,4- c ] pyridin-7-one ( 120 mg, 0.350 mmol) was dissolved in anhydrous acetonitrile (1.5 mL), and potassium carbonate (53.2 mg, 0.385 mmol) was added thereto, followed by stirring at room temperature for 30 minutes. 2-chloro-1- {4- (2,3-dimethylphenyl) piperazin-1-yl} ethanone (97.9 mg, 0.367 mmol) was added to the mixed solution and refluxed overnight. The reaction mixture was cooled to room temperature, extracted with methylene chloride, the organic layer was washed with water and the organic layer was dried over MgSO ₄ . The solid was filtered off, the solution was concentrated under reduced pressure, eluted by column chromatography (MeOH / CH ₂ Cl ₂ , 1:20), and purified to obtain 22.7 mg (0.048 mmol, 13.7%) of the title compound.

¹ H NMR (CDCl ₃ , 300 MHz) δ 7.24-7.17 (m, 4H), 7.09 (t, J = 7.6 Hz, 1H), 6.95 (d, J = 7.3 Hz, 1H), 6.87 (d, J = 7.8 Hz, 1H), 5.06 (s, 2H), 4.08 (s, 2H, NH ₂ ), 3.96 (t, J = 6.4 Hz, 2H), 3.80 (br, 4H), 2.89 (br, 4H), 2.77 (t, J = 6.4 Hz, 2H), 2.35 (s, 3H), 2.29 (s, 3H), 2.24 (s, 3H)

¹³ C NMR (CDCl ₃ , 75 MHz) δ 165.6, 161.3, 150.6, 142.2, 142.0, 140.3, 138.2, 136.0, 131.3, 129.5, 125.9, 125.7, 125.5, 116.8, 103.6, 52.3, 51.8, 50.5, 46.4, 42.7 , 21.0, 20.6, 19.6, 13.9

Example 2. 3-Amino-1-[{4- (2,3-dimethylphenyl) piperazin-1-yl} propanoyl] -6- N- ( p- tolyl) -4,5,6, 7-tetrahydro-1 H- pyrazolo [3,4- c ] pyridin-7-one (Compound No. 2)

1- Nt -butoxycarbonyl-3-amino-4,5,6,7-tetrahydro-6- N- ( p- tolyl) pyrazolo [3,4- c ] in the same manner as in Example 1 above Pyridin-7-one (120 mg, 0.350 mmol), potassium carbonate (53.2 mg, 0.385 mmol) and 3-chloro-1- (4- (2,3-dimethylphenyl) piperazin-1-yl) propane-1 -One (103.0 mg, 0.367 mmol) was reacted to give 80.9 mg (0.166 mmol, 47.5%) of the title compound as a white solid.

¹ H NMR (CDCl ₃ , 300 MHz) δ 7.22-7.16 (m, 4H), 7.07 (t, J = 7.7 Hz, 1H), 6.93 (d, J = 7.3 Hz, 1H), 6.84 (d, J = 7.9 Hz, 1H), 4.52 (s, 2H, NH ₂ ), 4.38 (t, J = 5.0 Hz, 2H), 3.91 (t, J = 6.4 Hz, 2H), 3.57 (br, 4H), 3.01 (t , J = 5.0 Hz, 2H), 2.81 (br, 4H), 2.72 (t, J = 6.4 Hz, 2H), 2.34 (s, 3H), 2.27 (s, 3H), 2.23 (s, 3H)

¹³ C NMR (CDCl ₃ , 75 MHz) δ 169.7, 161.6, 150.7, 142.3, 141.9, 140.4, 138.2, 135.8, 131.3, 129.5, 125.9, 125.6, 125.5, 116.7, 120.1, 52.1, 51.9, 45.9, 42.9, 42.4 , 33.4, 21.0, 20.6, 19.5, 13.8

Example 3. 3-Amino-1-{(4-phenylpiperazin-1-yl) propanoyl} -6- N- ( p- tolyl) -4,5,6,7-tetrahydro-1 H -pyrazolo [3,4 -c] pyridin-7-one (compound No. 3)

1- Nt -butoxycarbonyl-3-amino-4,5,6,7-tetrahydro-6- N- ( p- tolyl) pyrazolo [3,4- c ] in the same manner as in Example 1 above Pyridin-7-one (130 mg, 0.379 mmol), potassium carbonate (57.6 mg, 0.417 mmol) and 3-chloro-1- (4-phenylpiperazin-1-yl) propan-1-one (100.6 mg, 0.398 mmol) was reacted to give 77.8 mg (0.169 mmol, 44.7%) of the title compound in the liquid state.

¹ H NMR (CDCl ₃ , 300 MHz) δ 7.25 (t, J = 7.9 Hz, 2H), 7.15 (s, 4H), 6.88 (d, J = 8.8 Hz, 3H), 4.53 (s, 2H, NH ₂ ), 4.34 (t, J = 5.0 Hz, 2H), 3.83 (t, J = 6.4 Hz, 2H), 3.70 (br, 2H), 3.56 (br, 2H), 3.07 (br, 4H), 2.97 (t , J = 5.0 Hz, 2H), 2.66 (t, J = 6.4 Hz, 2H), 2.31 (s, 3H)

¹³ C NMR (CDCl ₃ , 75 MHz) δ 169.6, 161.6, 150.7, 142.1, 142.0, 140.3, 135.9, 129.5, 129.2, 125.5, 120.6, 116.6, 102.2, 51.9, 49.4, 49.2,45.3, 42.9, 41.8, 33.2 , 21.0, 19.4

Example 4. 3-Amino-1-[{4- (2,4-dimethylphenyl) piperazin-1-yl} propanoyl] -6- N- ( p- tolyl) -4,5,6, 7-tetrahydro-1 H- pyrazolo [3,4- c ] pyridin-7-one (Compound No. 4)

1- Nt -butoxycarbonyl-3-amino-4,5,6,7-tetrahydro-6- N- ( p- tolyl) pyrazolo [3,4- c ] in the same manner as in Example 1 above Pyridin-7-one (130 mg, 0.379 mmol), potassium carbonate (57.6 mg, 0.417 mmol) and 3-chloro-1- {4- (2,4-dimethylphenyl) piperazin-1-yl} propane-1 The reaction with -one (111.7 mg, 0.398 mmol) gave 102.8 mg (0.211 mmol, 55.7%) of the title compound as a white solid.

¹ H NMR (CDCl ₃ , 300 MHz) δ 7.21-7.15 (m, 4H), 7.01 (s, 1H), 6.96 (d, J = 8.2 Hz, 1H), 6.84 (d, J = 8.0 Hz, 1H) , 4.56 (s, 2H, NH ₂ ), 4.37 (t, J = 4.7 Hz, 2H), 3.89 (t, J = 6.4 Hz, 2H), 3.70 (br, 2H), 3.55 (br, 2H), 3.00 (t, J = 4.8 Hz, 2H), 2.79 (br, 4H), 2.70 (t, J = 6.4 Hz, 2H), 2.33 (s, 3H), 2.27 (s, 6H)

¹³ C NMR (CDCl ₃ , 75 MHz) δ 169.6, 161.6, 148.2, 142.2, 142.0, 140.4, 135.8, 133.3, 132.5, 131.9, 129.5, 127.1, 125.5, 119.0, 102.1, 51.9, 51.9, 51.7, 45.9, 42.9 , 42.4, 33.4, 21.0, 20.7, 19.5, 17.6

Example 5. 3-Amino-1-[{4-((benzo [ d ] [1,3] dioxol-5-yl) methyl) piperazin-1-yl} propanoyl] -6- N − ( p- tolyl) -4,5,6,7-tetrahydro-1 H- pyrazolo [3,4- c ] pyridin-7-one (Compound No. 5)

1- Nt -butoxycarbonyl-3-amino-4,5,6,7-tetrahydro-6- N- ( p- tolyl) pyrazolo [3,4-c] in the same manner as in Example 1 above Pyridin-7-one (130 mg, 0.379 mmol), potassium carbonate (57.6 mg, 0.417 mmol) and 1- [4-{(benzo [ d ] [1,3] dioxol-5-yl) methyl} piperazine -1-yl] -3-chloropropan-1-one (123.7 mg, 0.398 mmol) was reacted to give 101.7 mg (0.196 mmol, 51.9%) of the title compound as a white solid.

¹ H NMR (CDCl ₃ , 300 MHz) δ 7.19-7.11 (m, 4H), 6.79 (s, 1H), 6.72-6.66 (m, 2H), 5.90 (s, 2H), 4.53 (s, 2H, NH ₂ ), 4.29 (t, J = 4.8 Hz, 2H), 3.86 (t, J = 6.4 Hz, 2H), 3.53 (br, 2H), 3.38 (br, 2H), 3.36 (s, 2H), 2.90 ( t, J = 4.8 Hz, 2H), 2.66 (t, J = 6.3 Hz, 2H), 2.32 (br, 4H), 2.29 (s, 3H)

¹³ C NMR (CDCl ₃ , 75 MHz) δ 169.4, 161.6, 147.7, 146.7, 142.1, 142.0, 140.4, 135.8, 131.3, 129.5, 125.5, 122.1, 109.3, 107.9, 102.0, 100.9, 62.4, 52.6, 52.3, 51.9 , 45.2,42.8, 41.8, 33.3, 21.0, 19.5

Example 6. 3-Amino-1-[{4- (2,4,6-trimethylbenzyl) piperazin-1-yl} propanoyl] -6- N- ( p- tolyl) -4,5, 6,7-tetrahydro-1 H- pyrazolo [3,4- c ] pyridin-7-one (Compound No. 6)

3-Amino-5,6-dihydro-6- N- ( p- tolyl) -1H - pyrazolo [3,4- c ] pyridin-7 ( 4H ) -one in the same manner as in Example 1 above (108.2 mg, 0.446 mmol), potassium carbonate (92.4 mg, 0.669 mmol), 1- {4- (2,4,6-trimethylbenzyl) piperazin-1-yl} -3-chloropropan-1-one ( 151.3 mg, 0.490 mmol) was reacted to give 130.2 mg (0.253 mmol, 56.7%) of the title compound as a yellow solid.

¹ H NMR (CDCl ₃ , 300 MHz) δ 7.22-7.14 (m, 4H), 6.83 (s, 2H), 4.56 (s, 2H, NH ₂ ), 4.32 (t, J = 4.8 Hz, 2H), 3.89 (t, J = 6.4 Hz, 2H), 3.49 (br, 2H), 3.44 (s, 2H), 3.34 (br, 2H), 2.93 (t, J = 4.9 Hz, 2H), 2.69 (t, J = 6.4 Hz, 2H), 2.38 (br, 4H), 2.32 (s, 9H), 2.26 (s, 3H)

¹³ C NMR (CDCl ₃ , 75 MHz) δ 169.3, 161.6, 142.1, 142.0, 140.4, 138.0, 136.6, 135.8, 131.0, 129.6, 129.5, 128.9, 125.5, 125.3, 102.0, 55.6, 52.4, 52.1, 51.9, 45.6 , 42.8, 42.2,33.4, 21.0, 20.9, 20.0, 19.5

Example 7. 3-Amino-1-[{4- (4- t -butylbenzyl) piperazin-1-yl} propanoyl] -6- N- ( p- tolyl) -4,5,6, 7-tetrahydro-1 H- pyrazolo [3,4- c ] pyridin-7-one (Compound No. 7)

3-Amino-5,6-dihydro-6- N- ( p- tolyl) -1H - pyrazolo [3,4- c ] pyridin-7 ( 4H ) -one in the same manner as in Example 1 above (100 mg, 0.412 mmol), potassium carbonate (85.4 mg, 0.618 mmol) and 1- {4- (4 -t- butylbenzyl) piperazin-1-yl} -3-chloropropan-1-one (146.2 mg , 0.453 mmol) to give 109.8 mg (0.207 mmol, 50.4%) of the title compound as a yellow solid.

¹ H NMR (CDCl ₃ , 300 MHz) δ 7.33 (d, J = 8.2 Hz, 2H), 7.22-7.14 (m, 6H), 4.54 (s, 2H, NH ₂ ), 4.32 (t, J = 4.8 Hz , 2H), 3.89 (t, J = 6.4 Hz, 2H), 3.56 (br, 2H), 3.45 (s, 2H), 3.40 (br, 2H), 2.92 (t, J = 4.8 Hz, 2H), 2.69 (t, J = 6.4 Hz, 2H), 2.37 (br, 4H), 2.32 (s, 3H), 1.31 (s, 9H)

¹³ C NMR (CDCl ₃ , 75 MHz) δ 169.4, 161.6, 150.2, 142.2, 142.0, 140.4, 135.8, 134.3, 129.5, 128.8, 125.5, 125.2, 102.0, 62.4, 52.7, 52.5, 51.9, 45.3, 42.8, 41.8 , 34.5, 33.4, 31.4, 21.0, 19.5.

Example 8. 3-amino-1-[{4- (2,3,4-trimethoxybenzyl) piperazin-1-yl} propanoyl] -6- N- ( p- tolyl) -4, 5,6,7-tetrahydro-1 H- pyrazolo [3,4- c ] pyridin-7-one (Compound No. 8)

3-Amino-5,6-dihydro-6- N- ( p- tolyl) -1H - pyrazolo [3,4- c ] pyridin-7 ( 4H ) -one in the same manner as in Example 1 above (126.8 mg, 0.523 mmol), potassium carbonate (108.3 mg, 0.784 mmol) and 1- {4- (2,3,4-trimethoxybenzyl) piperazin-1-yl} -3-chloropropane-1- (205.1 mg, 0.575 mmol) were reacted to give 158.4 mg (0.281 mmol, 53.8%) of the title compound as a white solid.

¹ H NMR (CDCl ₃ , 300 MHz) δ 7.15-7.07 (m, 4H), 6.89 (d, J = 8.5 Hz, 1H), 6.57 (d, J = 8.5 Hz, 1H), 4.53 (s, 2H, NH ₂ ), 4.25 (t, J = 4.7 Hz, 2H), 3.82 (t, J = 6.9 Hz, 2H), 3.80 (s, 6H), 3.78 (s, 3H), 3.48 (br, 2H), 3.40 (s, 2H), 3.32 (br, 2H), 2.86 (t, J = 4.6 Hz, 2H), 2.63 (t, J = 6.4 Hz, 2H), 2.34 (br, 4H), 2.26 (s, 3H)

¹³ C NMR (CDCl ₃ , 75 MHz) δ 169.3, 161.6, 153.0, 152.5, 142.2, 142.0, 140.3, 135.7, 129.4, 125.5, 125.1, 123.1, 107.0, 102.0, 61.1, 60.7, 56.2, 55.9, 52.6, 52.3 , 51.9, 45.3, 42.8, 41.8, 33.2, 20.9, 19.4

Example 9. 3-Amino-1-[{4- (4-chlorophenyl) piperazin-1-yl} propanoyl] -6- N- ( p- tolyl) -4,5,6,7- Tetrahydro-1 H- pyrazolo [3,4- c ] pyridin-7-one (Compound No. 9)

3-Amino-5,6-dihydro-6- N- ( p- tolyl) -1H - pyrazolo [3,4- c ] pyridin-7 ( 4H ) -one in the same manner as in Example 1 above (101 mg, 0.416 mmol), potassium carbonate (86.2 mg, 0.624 mmol) and 3-chloro-1- {4- (4-chlorophenyl) piperazin-1-yl} propan-1-one (131.2 mg, 0.457 mmol) was reacted to give 117.9 mg (0.239 mmol, 57.5%) of the title compound as a white solid.

¹ H NMR (CDCl ₃ , 300 MHz) δ 7.18 (d, J = 8.8 Hz, 2H), 7.14 (s, 4H), 6.78 (d, J = 8.8 Hz, 2H), 4.47 (s, 2H, NH ₂ ), 4.34 (t, J = 4.8 Hz, 2H), 3.83 (t, J = 6.3 Hz, 2H), 3.68 (br, 2H), 3.53 (br, 2H), 3.02 (br, 4H), 2.97 (t , J = 4.8 Hz, 2H), 2.66 (t, J = 6.3 Hz, 2H), 2.30 (s, 3H)

¹³ C NMR (CDCl ₃ , 75 MHz) δ 169.6, 161.5, 149.4, 142.2, 141.9, 140.4, 135.8, 129.5, 129.1, 125.5, 125.3, 117.7, 102.2, 51.9, 49.3, 49.1, 45.1, 42.9, 41.6, 33.2 , 21.0, 19.5

Example 10. 3-Amino-1-[{4- (4-methoxyphenyl) piperazin-1-yl} propanoyl] -6- N- ( p- tolyl) -4,5,6,7 -Tetrahydro-1 H- pyrazolo [3,4- c ] pyridin-7-one (Compound No. 10)

3-Amino-5,6-dihydro-6- N- ( p- tolyl) -1H - pyrazolo [3,4- c ] pyridin-7 ( 4H ) -one in the same manner as in Example 1 above (100 mg, 0.412 mmol), potassium carbonate (85.4 mg, 0.618 mmol) and 3-chloro-1- {4- (4-methoxyphenyl) piperazin-1-yl} propan-1-one (128.1 mg, 0.453 mmol) was reacted to give 154.9 mg (0.317 mmol, 76.9%) of the title compound as a white solid.

¹ H NMR (CDCl ₃ , 300 MHz) δ 7.17-7.10 (m, 4H), 6.85-6.78 (m, 4H), 4.50 (s, 2H, NH ₂ ), 4.33 (t, J = 4.5 Hz, 2H) , 3.82 (t, J = 6.4 Hz, 2H), 3.72 (s, 3H), 3.68 (br, 2H), 3.52 (br, 2H), 2.94 (br, 6H), 2.64 (t, J = 6.3 Hz, 2H), 2.29 (s, 3H)

¹³ C NMR (CDCl ₃ , 75 MHz) δ 169.5, 161.6, 154.3, 145.0, 142.2, 141.9, 140.4, 135.7, 129.4, 125.5, 118.8, 114.5, 102.1, 55.5, 51.9, 50.9, 50.7, 45.4, 42.9, 41.9 , 33.2, 21.0, 19.4

Example 11. 3-Amino-1-{( N -benzyl) piperazin-1-yl} -6- N- ( p -methoxyphenyl) -4,5,6,7-tetrahydro-pyrazolo [ 3,4- c ] pyridin-7-one (Compound No. 11)

1- Nt -butoxycarbonyl-3-amido-4,5,6,7-tetrahydro-6- N- (4-methoxyphenyl) pyrazolo [3,4 in the same manner as in Example 1 above. c ] pyridin-7-one (173 mg, 0.482 mmol), potassium carbonate (73.2 mg, 0.530 mmol) and 1-{( N- benzyl) piperazin-1-yl} -3-chloropropan-1-one (100 mg, 0.506 mmol) was reacted to obtain 20 mg (0.047 mmol, 9.9%) of the target compound in the form of a white solid.

¹ H NMR (CDCl ₃ , 300 MHz) δ 7.33-7.25 (m, 3H), 7.21 (d, J = 7.7 Hz, 2H), 7.10 (d, J = 8.7 Hz, 2H), 6.91 (d, J = 8.7 Hz, 2H), 4.68 (t, J = 6.5 Hz, 2H), 4.42 (d, J = 5.6 Hz, 2H), 3.90 (t, J = 6.6 Hz, 2H), 3.82 (s, 3H), 3.57 (s, 2H, NH ₂ ), 2.84 (t, J = 6.5 Hz, 2H), 2.74 (t, J = 6.6 Hz, 2H)

¹³ C NMR (CDCl ₃ , 75 MHz) δ 170.2, 159.1, 158.1, 148.8, 138.4, 134.6, 131.4, 128.5, 127.7, 127.2, 126.9, 114.3, 107.5, 55.5, 51.9, 46.2,43.4, 37.5, 19.4

Example 12. 3-Amino-1-[{4- (2,4-dimethylphenyl) piperazin-1-yl} propanoyl] -6- N- ( p -methoxyphenyl) -4,5, 6,7-tetrahydro-1 H- pyrazolo [3,4- c ] pyridin-7-one (Compound No. 12)

T- butyl 3-amino-4,5,6,7-tetrahydro-6- N- (4-methoxyphenyl) -7-oxopyrazolo [3,4- c ] in the same manner as in Example 1 above Pyridine-1-carboxyester (161.3 mg, 0.45 mmol), potassium carbonate (68.4 mg, 0.495 mmol) and 3-chloro-1- {4- (2,4-dimethylphenyl) piperazin-1-yl} propane The reaction of -1-one (132.5 mg, 0.472 mmol) gave 70.2 mg (0.139 mmol, 31.0%) of the title compound in the form of a white solid.

¹ H NMR (CDCl ₃ , 300 MHz) δ 7.21 (d, J = 8.7 Hz, 2H), 7.00 (s, 1H), 6.95 (d, J = 8.2 Hz, 1H), 6.88 (d, J = 8.7 Hz , 2H), 6.84 (d, J = 8.1 Hz, 1H), 4.56 (s, 2H, NH ₂ ), 4.36 (t, J = 4.7 Hz, 2H), 3.87 (t, J = 6.3 Hz, 2H), 3.78 (s, 3H), 3.69 (br, 2H), 3.54 (br, 2H), 2.99 (t, J = 4.7 Hz, 2H), 2.78 (br, 4H), 2.70 (t, J = 6.3 Hz, 2H 2.26 (s, 6H)

¹³ C NMR (CDCl ₃ , 75 MHz) δ 169.6, 161.8, 157.7, 148.2, 142.2, 142.0, 135.9, 133.0, 132.5, 131.9, 127.1, 127.0, 119.0, 114.2, 102.0, 55.4, 52.2, 51.9, 51.7, 45.9 , 42.8, 42.4, 33.4, 20.7, 19.5, 17.6

Example 13. 3-amino-1-[{4- (2,4,6-trimethylbenzyl) piperazin-1-yl} propanoyl] -6- N- ( p -methoxyphenyl) -4, 5,6,7-tetrahydro-1 H- pyrazolo [3,4- c ] pyridin-7-one (Compound No. 13)

3-Amino-5,6-dihydro-6- N- (4-methoxyphenyl) -1 H- pyrazolo [3,4- c ] pyridine-7 ( 4H ) in the same manner as in Example 1 -One (120 mg, 0.464 mmol), potassium carbonate (96.2 mg, 0.696 mmol) and 1- {4- (2,4,6-trimethylbenzyl) piperazin-1-yl} -3-chloropropane-1- Warmed (157.5 mg, 0.510 mmol) gave 81.2 mg (0.153 mmol, 32.9%) of the title compound as a white solid.

¹ H NMR (CDCl ₃ , 300 MHz) δ 7.23 (d, J = 8.7 Hz, 2H), 6.88 (d, J = 8.7 Hz, 2H), 6.83 (s, 2H), 4.54 (s, 2H, NH ₂ ), 4.32 (t, J = 4.7 Hz, 2H), 3.87 (t, J = 6.3 Hz, 2H), 3.78 (s, 3H), 3.49 (br, 2H), 3.44 (s, 2H), 3.33 (br , 2H), 2.93 (t, J = 4.9 Hz, 2H), 2.69 (t, J = 6.2 Hz, 2H), 2.38 (br, 4H), 2.32 (s, 6H), 2.25 (s, 3H)

¹³ C NMR (CDCl ₃ , 75 MHz) δ 169.3, 161.7, 157.7, 142.2, 141.9, 138.0, 136.6, 135.9, 131.0, 128.9, 126.9, 126.8, 114.3, 114.2, 101.9, 55.6, 55.4, 52.4, 52.2, 52.1 , 45.5, 42.7, 42.1,33.4, 20.9, 20.0, 19.5

Example 14. 3-amino-1-[{4- (2,3,4-trimethoxybenzyl) piperazin-1-yl} propanoyl] -6- N- ( p -methoxyphenyl)- 4,5,6,7-tetrahydro-1 H- pyrazolo [3,4- c ] pyridin-7-one (Compound No. 14)

3-Amino-5,6-dihydro-6- N- (4-methoxyphenyl) -1 H- pyrazolo [3,4- c ] pyridine-7 ( 4H ) in the same manner as in Example 1 -One (129.8 mg, 0.502 mmol), potassium carbonate (104.0 mg, 0.753 mmol) and 1- {4- (2,3,4-trimethoxybenzyl) piperazin-1-yl} -3-chloropropane- 1-one (196.9 mg, 0.552 mmol) was reacted to give 136.1 mg (0.235 mmol, 46.8%) of the title compound as a white solid.

¹ H NMR (CDCl ₃ , 300 MHz) δ 7.18 (d, J = 8.8 Hz, 2H), 6.91 (d, J = 8.5 Hz, 1H), 6.84 (d, J = 8.8 Hz, 2H), 6.59 (d , J = 8.5 Hz, 1H), 4.51 (s, 2H, NH ₂ ), 4.28 (t, J = 4.5 Hz, 2H), 3.85 (br, 2H), 3.82 (s, 6H), 3.80 (s, 3H ), 3.73 (s, 3H), 3.51 (br, 2H), 3.42 (s, 2H), 3.36 (br, 2H), 2.89 (t, J = 4.5 Hz, 2H), 2.65 (t, J = 6.3 Hz , 2H), 2.36 (br, 4H)

¹³ C NMR (CDCl ₃ , 75 MHz) δ 169.3, 161.7, 157.6, 153.0, 152.5, 142.2, 142.1, 141.9, 135.9, 126.9, 125.0, 123.2, 114.2, 107.0, 101.9, 61.1, 60.7, 56.3, 55.9, 55.4 , 52.6, 52.3, 52.1, 45.3, 42.8, 41.9, 33.3, 19.4

Example 15. 3-Amino-1-[{4- (2,4-dimethylphenyl) piperazin-1-yl} propanoyl] -6- N -phenyl-4,5,6,7-tetrahydro -Pyrazolo [3,4- c ] pyridin-7-one (Compound No. 15)

3-Amino-5,6-dihydro-6-phenyl-1 H- pyrazolo [3,4- c ] pyridin-7 ( 4H ) -one (120 mg, 0.525 mmol) in the same manner as in Example 1 above ), Potassium carbonate (108.7 mg, 0.787 mmol) and 3-chloro-1- {4- (2,4-dimethylphenyl) piperazin-1-yl} propan-1-one (162 mg, 0.577 mmol) Thus, 152 mg (0.321 mmol, 61.2%) of the title compound were obtained.

¹ H NMR (CDCl ₃ , 300 MHz) δ 7.38-7.27 (m, 4H), 7.19 (t, J = 6.7 Hz, 1H), 7.01 (s, 1H), 6.95 (d, J = 8.0 Hz, 1H) , 6.84 (d, J = 8.0 Hz, 1H), 4.58 (s, 2H, NH ₂ ), 4.37 (t, J = 4.5 Hz, 2H), 3.91 (t, J = 6.3 Hz, 2H), 3.69 (br , 2H), 3.53 (br, 2H), 2.99 (t, J = 4.5 Hz, 2H), 2.78 (br, 4H), 2.70 (t, J = 6.3 Hz, 2H), 2.26 (s, 6H)

¹³ C NMR (CDCl ₃ , 75 MHz) δ 169.6, 161.6, 148.2, 143.0, 142.2, 142.1, 133.3, 132.5, 131.9, 128.8, 127.1, 126.0, 125.7, 119.0, 102.1, 51.9, 51.7, 45.9, 42.9, 42.4 , 33.4, 20.7, 19.5, 17.6

Example 16. 3-Amino-1-[{4- (2,4,6-trimethylbenzyl) piperazin-1-yl} propanoyl] -6- N -phenyl-4,5,6,7- Tetrahydropyrazolo [3,4- c ] pyridin-7-one (Compound No. 16)

3-Amino-5,6-dihydro-6-phenyl-1 H- pyrazolo [3,4- c ] pyridin-7 ( 4H ) -one (120 mg, 0.525 mmol) in the same manner as in Example 1 above ), Potassium carbonate (108.7 mg, 0.787 mmol) and 3-chloro-1- {4- (2,4,6-trimethylbenzyl) piperazin-1-yl} propan-1-one (178.2 mg, 0.577 mmol) Was reacted to obtain 92.7 mg (0.185 mmol, 35.3%) of the title compound.

¹ H NMR (CDCl ₃ , 300 MHz) δ 7.39-7.31 (m, 4H), 7.20 (t, J = 6.6 Hz, 1H), 6.84 (s, 2H), 4.56 (s, 2H, NH ₂ ), 4.34 (t, J = 4.7 Hz, 2H), 3.93 (t, J = 6.3 Hz, 2H), 3.50 (br, 2H), 3.44 (s, 2H), 3.34 (br, 2H), 2.94 (t, J = 4.7 Hz, 2H), 2.71 (t, J = 6.3 Hz, 2H), 2.38 (br, 4H), 2.32 (s, 6H), 2.26 (s, 3H)

¹³ C NMR (CDCl ₃ , 75 MHz) δ 169.3, 161.6, 143.0, 142.1, 142.0, 138.0, 136.6, 131.0, 129.0, 128.9, 126.0, 125.6, 125.5, 102.0, 55.6, 52.4, 52.1, 51.9, 45.5, 42.8 , 42.2, 33.4, 20.9, 20.0, 19.5

Example 17. 3-Amino-1-[{4- (2,4-dimethylphenyl) piperazin-1-yl} propanoyl] -6- N -benzyl-4,5,6,7-tetrahydro -1 H - pyrazolo [3,4- c] pyridin-7-one (compound No. 17)

T- butyl-3-amino-6-benzyl-4,5,6,7-tetrahydro-7-oxapyrazolo [3,4- c ] pyridine-1-carboxyl dissolved in anhydrous acetonitrile (1.9 mL) Potassium carbonate (68.2 mg, 0.494 mmol) was added to a solution of rate (154 mg, 0.449 mmol) and stirred at room temperature for 30 minutes. 3-chloro-1- {4- (2,4-dimethylphenyl) piperazin-1-yl} propan-1-one (132.2 mg, 0.471 mmol) was added to the reaction mixture and reacted overnight. 25.4 mg (0.052 mmol, 11.6%) was obtained.

¹ H NMR (CDCl ₃ , 300 MHz) δ 7.32-7.27 (m, 5H), 7.02 (s, 1H), 6.95 (br, 1H), 6.85 (d, J = 8.0 Hz, 1H), 4.75 (s, 2H), 4.45 (s, 2H, NH ₂ ), 4.36 (t, J = 5.0 Hz, 2H), 3.69 (t, J = 4.5 Hz, 2H), 3.56 (t, J = 4.7 Hz, 2H), 3.43 (t, J = 6.6 Hz, 2H), 3.01 (t, J = 4.9 Hz, 2H), 2.84-2.77 (m, 4H), 2.55 (t, J = 6.6 Hz, 2H), 2.27 (s, 6H)

¹³ C NMR (CDCl ₃ , 75 MHz) δ 169.6, 162.1, 148.2, 141.9, 141.8, 137.7, 133.3, 132.6, 131.9, 128.5, 128.2, 128.1, 127.3, 127.1, 119.0, 101.5, 51.9, 51.7, 49.4, 47.3 , 45.9, 42.7, 42.4, 33.4, 20.7, 18.9, 17.6

Example 18. 3-Amino-1-{(4-phenylpiperazin-1-yl) propanoyl} -6- N- { p- (dimethylamino) phenyl} -4,5,6,7- Tetrahydro-1 H- pyrazolo [3,4- c ] pyridin-7-one (Compound No. 18)

In the same manner as Example 1 3-Amino -6- N - (4- (dimethylamino) phenyl) -5,6-dihydro -1 H- pyrazolo [3,4 -c] pyridine-7 ( 4 H ) -one (110 mg, 0.405 mmol), potassium carbonate (83.9 mg, 0.607 mmol) and 3-chloro-1- (4-phenylpiperazin-1-yl) propan-1-one (112.4 mg, 0.445 mmol) was reacted to give 15.4 mg (0.031 mmol, 7.8%) of the title compound in the solid state.

¹ H NMR (CDCl ₃ , 300 MHz) δ 7.29 (t, J = 7.8 Hz, 2H), 7.17 (d, J = 8.8 Hz, 2H), 6.93-6.90 (m, 3H), 6.73 (d, J = 8.8 Hz, 2H), 4.46 (s, 2H, NH ₂ ), 4.38 (t, J = 4.8 Hz, 2H), 3.86 (t, J = 6.4 Hz, 2H), 3.74 (br, 2H), 3.59 (br , 2H), 3.12 (br, 4H), 3.02 (t, J = 4.8 Hz, 2H), 2.94 (s, 6H), 2.70 (t, J = 6.4 Hz, 2H)

¹³ C NMR (CDCl ₃ , 75 MHz) δ 169.7, 161.7, 150.7, 149.1, 142.5, 141.7, 132.3, 129.3, 126.5, 120.7, 116.7, 112.9, 102.0, 52.2,49.4, 49.3, 45.3, 42.7, 41.8, 40.8 , 33.4, 19.5

Example 19. 3-Amino-1-[{4- (4-methoxyphenyl) piperazin-1-yl} propanoyl] -6- N- { p- (dimethylamino) phenyl} -4, 5,6,7-tetrahydro-1 H- pyrazolo [3,4- c ] pyridin-7-one (Compound No. 19)

In the same manner as Example 1 3-Amino -6- N - (4- (dimethylamino) phenyl) -5,6-dihydro -1 H- pyrazolo [3,4 -c] pyridine-7 ( 4 H ) -one (105 mg, 0.387 mmol), potassium carbonate (80.1 mg, 0.580 mmol) and 3-chloro-1- {4- (4-methoxyphenyl) piperazin-1-yl} propane-1- Warm (120.1 mg, 0.425 mmol) was reacted to give 22.2 mg (0.043 mmol, 11.1%) of the title compound.

¹ H NMR (CDCl ₃ , 300 MHz) δ 7.16 (d, J = 8.9 Hz, 2H), 6.90-6.82 (m, 4H), 6.72 (d, J = 8.9 Hz, 2H), 4.43 (s, 2H, NH ₂ ), 4.38 (t, J = 4.9 Hz, 2H), 3.86 (t, J = 6.4 Hz, 2H), 3.77 (s, 3H), 3.73 (br, 2H), 3.58 (br, 2H), 3.00 (br, 6H), 2.93 (s, 6H), 2.69 (t, J = 6.4 Hz, 2H)

¹³ C NMR (CDCl ₃ , 75 MHz) δ 169.6, 161.7, 154.4, 149.0, 145.0, 142.5, 141.6, 132.4, 126.5, 118.9, 114.5, 112.9, 102.0, 55.5, 52.2, 50.9, 50.8, 45.5, 42.8, 41.9 , 40.8, 33.3, 19.5

Example 20. 3-Amino-1-{(4-phenylpiperazin-1-yl) propanoyl} -6- N -{(furan-2-yl) methyl} -4,5,6,7- Tetrahydro-1 H- pyrazolo [3,4- c ] pyridin-7-one (Compound No. 20)

In the same manner as Example 1 3-Amino -6- N - ((furan-2-yl) methyl) -5,6-dihydro -1 H- pyrazolo [3,4 -c] pyridine-7 ( 4 H ) -one (104 mg, 0.447 mmol), potassium carbonate (92.6 mg, 0.670 mmol) and 3-chloro-1- (4-phenylpiperazin-1-yl) propan-1-one (124.1 mg, 0.491 mmol) was reacted to give 192.8 mg (0.429 mmol, 96.2%) of the title compound in the solid state.

¹ H NMR (CDCl ₃ , 300 MHz) δ 7.30-7.23 (m, 3H), 6.91-6.86 (m, 3H), 6.29-6.25 (m, 2H), 4.68 (s, 2H), 4.41 (s, 2H , NH ₂ ), 4.33 (t, J = 4.9 Hz, 2H), 3.69 (br, 2H), 3.26 (br, 2H), 3.48 (t, J = 6.5 Hz, 2H), 3.08 (br, 4H), 2.97 (t, J = 4.9 Hz, 2H), 2.54 (t, J = 6.5 Hz, 2H)

¹³ C NMR (CDCl ₃ , 75 MHz) δ 169.5, 161.8, 151.1, 150.7, 142.0, 141.9, 141.6, 129.2, 120.5, 116.5, 110.3, 108.1, 101.8, 49.3, 49.1, 47.7, 45.2,42.8, 42.3, 41.7 , 33.1, 18.7

Example 21. 3-amino-1-[{4- (4-methoxyphenyl) piperazin-1-yl} propanoyl] -6- N -{(furan-2-yl) methyl} -4, 5,6,7-tetrahydropyrazolo [3,4- c ] pyridin-7-one (Compound No. 21)

In the same manner as Example 1 3-Amino -6- N - ((furan-2-yl) methyl) -5,6-dihydro -1 H- pyrazolo [3,4 -c] pyridine-7 ( 4 H ) -one (100 mg, 0.430 mmol), potassium carbonate (89.1 mg, 0.645 mmol) and 3-chloro-1- {4- (4-methoxyphenyl) piperazin-1-yl} propane-1- Warm (133.7 mg, 0.473 mmol) was reacted to give 191.5 mg (0.400 mmol, 93.1%) of the title compound in the solid state.

¹ H NMR (CDCl ₃ , 300 MHz) δ 7.23 (s, 1H), 6.79-6.72 (m, 4H), 6.21-6.17 (m, 2H), 4.60 (s, 2H), 4.48 (s, 2H, NH ₂ ), 4.25 (t, J = 4.7 Hz, 2H), 3.67 (s, 3H), 3.61 (br, 2H), 3.46 (br, 2H), 3.40 (t, J = 6.6 Hz, 2H), 2.88 ( br, 6H), 2.48 (t, J = 6.5 Hz, 2H)

¹³ C NMR (CDCl ₃ , 75 MHz) δ 169.4, 161.8, 154.3, 151.1, 145.0, 142.0, 141.9, 141.6, 118.8, 114.4, 110.3, 108.1, 101.7, 55.4, 50.8, 50.6, 47.6, 45.3, 42.8, 42.3 , 41.8, 33.2, 18.8

Example 22. 3-Amino-1- {4- (2-pyridinylpiperazin-1-yl) propanoyl} -6- Np -tolyl-4,5,6,7-tetrahydro-1 H- Pyrazolo [3,4- c ] pyridin-7-one (Compound No. 22)

3-Amino-5,6-dihydro-6- N- ( p- tolyl) -pyrazolo [3,4- c ] pyridin-7 ( 4H ) -one (105 mg) in the same manner as in Example 1 above , 0.433 mmol), potassium carbonate (89.7 mg, 0.649 mmol) and 3-chloro-1- {4- (pyridin-2-yl) piperazin-1-yl} propan-1-one (120.7 mg, 0.476 mmol) Reaction was carried out to give 159.6 mg (0.347 mmol, 80.2%) of the title compound in the solid state.

¹ H NMR (CDCl ₃ , 300 MHz) δ 8.11 (d, J = 3.3 Hz, 1H), 7.41 (t, J = 7.8 Hz, 1H), 7.13-7.06 (m, 4H), 6.61-6.54 (m, 2H), 4.51 (s, 2H, NH ₂ ), 4.30 (t, J = 4.8 Hz, 2H), 3.78 (t, J = 6.4 Hz, 2H), 3.62 (t, J = 4.7 Hz, 2H), 3.44 (br, 6H), 2.92 (t, J = 4.8 Hz, 2H), 2.61 (t, J = 6.4 Hz, 2H), 2.25 (s, 3H)

¹³ C NMR (CDCl ₃ , 75 MHz) δ 169.7, 161.6, 158.8, 147.9, 142.1, 142.0, 140.4, 137.6, 135.7, 129.4, 125.5, 113.9, 107.2, 102.1, 51.9, 44.9, 44.8, 44.7, 42.8, 41.4 , 33.3, 20.9, 19.4

Example 23. 3-Amino-1- {4- (2-pyridinylpiperazin-1-yl) propanoyl} -6- N -{(furan-2-yl) methyl} -4,5,6 , 7-tetrahydro-1 H -pyrazolo [3,4- c ] pyridin-7-one (Compound No. 23)

In the same manner as Example 1 3-Amino -6- N - ((furan-2-yl) methyl) -5,6-dihydro -1 H- pyrazolo [3,4 -c] pyridine-7 ( 4 H ) -one (103 mg, 0.443 mmol), potassium carbonate (91.7 mg, 0.664 mmol) and 3-chloro-1- {4- (pyridin-2-yl) piperazin-1-yl} propane-1- Warmed (123.5 mg, 0.487 mmol) to give 191.6 mg (0.426 mmol, 96.2%) of the title compound in the solid state.

¹ H NMR (CDCl ₃ , 300 MHz) δ 8.07 (d, J = 6.0 Hz, 1H), 7.39 (t, J = 7.8 Hz, 1H), 7.21 (s, 1H), 6.57-6.51 (m, 2H) , 6.19-6.15 (m, 2H), 4.59 (s, 2H), 4.46 (s, 2H, NH ₂ ), 4.24 (t, J = 4.9 Hz, 2H), 3.57 (t, J = 5.0 Hz, 2H) , 3.42-3.34 (m, 8H), 2.88 (t, J = 4.9 Hz, 2H), 2.46 (t, J = 6.5 Hz, 2H)

¹³ C NMR (CDCl ₃ , 75 MHz) δ 169.6, 161.8, 158.7, 151.1, 147.8, 142.0, 141.9, 141.7, 137.6, 113.9, 110.3, 108.1, 107.1, 101.7, 47.6, 44.9, 44.8, 44.7, 42.7, 42.2 , 41.4, 33.3, 18.8

Example 24. 3-Amino-1-[{4- (4-trifluorofluoromethylphenyl) piperazin-1-yl} propanoyl] -6- N- ( p -tolyl) -4,5,6 , 7-tetrahydro-1 H -pyrazolo [3,4- c ] pyridin-7-one (Compound No. 24)

3-Amino-5,6-dihydro-6- N- ( p- tolyl) -1H - pyrazolo [3,4- c ] pyridin-7 ( 4H ) -one in the same manner as in Example 1 above (95 mg, 0.392 mmol), potassium carbonate (81.2 mg, 0.588 mmol) and 3-chloro-1- [4- {4- (trifuluromethyl) phenyl} piperazin-1-yl] propane-1- Warm (138.2 mg, 0.431 mmol) was obtained to give 176.3 mg (0.334 mmol, 85.4%) of the title compound in the solid state.

¹ H NMR (CDCl ₃ , 300 MHz) δ 7.48 (d, J = 8.6 Hz, 2H), 7.15 (s, 4H), 6.89 (d, J = 8.6 Hz, 2H), 4.43 (s, 2H, NH ₂ ), 4.37 (t, J = 5.0 Hz, 2H), 3.86 (t, J = 6.4 Hz, 2H), 3.72 (t, J = 4.9 Hz, 2H), 3.58 (t, J = 4.9 Hz, 2H), 3.20 (br, 4H), 3.00 (t, J = 5.0 Hz, 2H), 2.68 (t, J = 6.4 Hz, 2H), 2.32 (s, 3H)

¹³ C NMR (CDCl ₃ , 75 MHz) δ 169.7, 161.5, 152.7, 142.3, 141.8, 140.4, 135.8, 129.5, 126.5, 126.4, 125.5, 114.9, 120.2, 51.9, 48.0, 47.9, 44.9, 42.8, 41.4, 33.2 , 20.9, 19.5

Example 25. 3-Amino-1-[{4- (2,4-dimethylphenyl) piperazin-1-yl} propanoyl] -6- N -{(furan-2-yl) methyl} -4 , 5,6,7-tetrahydro-1 H -pyrazolo [3,4- c ] pyridin-7-one (Compound No. 25)

In the same manner as Example 1 3-Amino -6- N - ((furan-2-yl) methyl) -5,6-dihydro -1 H- pyrazolo [3,4 -c] pyridine-7 ( 4 H ) -one (98 mg, 0.422 mmol), potassium carbonate (87.4 mg, 0.633 mmol) and 3-chloro-1- {4- (2,4-dimethylphenyl) piperazin-1-yl} propane-1 -One (130.2 mg, 0.464 mmol) was reacted to give 187.7 mg (0.393 mmol, 93.3%) of the title compound in the solid state.

¹ H NMR (CDCl ₃ , 300 MHz) δ 7.33 (s, 1H), 7.01 (s, 1H), 6.97 (d, J = 8.0 Hz, 1H), 6.85 (d, J = 8.0 Hz, 1H), 6.32 -6.28 (m, 2H), 4.73 (s, 2H), 4.42 (s, 2H, NH ₂ ), 4.35 (t, J = 5.0 Hz, 2H), 3.69 (t, J = 4.5 Hz, 2H), 3.56 -3.49 (m, 4H), 2.99 (t, J = 5.0 Hz, 2H), 2.83-2.77 (m, 4H), 2.59 (t, J = 6.6 Hz, 2H), 2.27 (s, 6H)

¹³ C NMR (CDCl ₃ , 75 MHz) δ 169.5, 161.9, 151.2, 148.2, 142.0, 141.6, 133.1, 132.4, 131.8, 127.1, 119.0, 110.3, 108.1, 101.7, 51.8, 51.6, 47.7, 45.8, 42.8, 42.3 , 33.3, 20.7, 18.8, 17.6

Example 26. 3-Amino-1-[{4- (2,4,5-trimethylphenyl) piperazin-1-yl} propanoyl] -6- N- ( p -tolyl) -4,5, 6,7-tetrahydro-1 H -pyrazolo [3,4- c ] pyridin-7-one (Compound No. 26)

3-Amino-5,6-dihydro-6- N- ( p- tolyl) -1H - pyrazolo [3,4- c ] pyridin-7 ( 4H ) -one in the same manner as in Example 1 above (97 mg, 0.4 mmol), potassium carbonate (82.9 mg, 0.6 mmol) and 3-chloro-1- {4- (2,4,5-trimethylphenyl) piperazin-1-yl} propan-1-one ( 129.7 mg, 0.44 mmol) was reacted to give 107.5 mg (0.214 mmol, 53.7%) of the title compound in the solid state.

¹ H NMR (CDCl ₃ , 300 MHz) δ 7.22-7.15 (m, 4H), 6.96 (s, 1H), 6.74 (s, 1H), 4.55 (s, 2H, NH ₂ ), 4.37 (t, J = 4.9 Hz, 2H), 3.91 (t, J = 6.4 Hz, 2H), 3.70 (br, 2H), 3.55 (br, 2H), 3.00 (t, J = 4.9 Hz, 2H), 2.83-2.78 (m, 4H), 2.72 (t, J = 6.4 Hz, 2H), 2.33 (s, 3H), 2.24 (s, 3H), 2.19 (s, 3H), 2.18 (s, 3H)

¹³ C NMR (CDCl ₃ , 75 MHz) δ 169.6, 161.6, 148.4, 142.2, 141.9, 140.4, 135.8, 134.5, 132.4, 131.8, 129.8, 129.5, 125.5, 120.5, 102.1, 51.9, 51.7, 50.5, 45.9, 42.8 , 42.5, 33.4, 21.0, 19.5, 18.9, 17.1

Example 27. 3-Amino-1-[{4- (4- t -butylphenyl) piperazin-1-yl} propanoyl] -6- N- ( p -tolyl) -4,5,6, 7-tetrahydro-1 H -pyrazolo [3,4- c ] pyridin-7-one (Compound No. 27)

3-Amino-5,6-dihydro-6- N- ( p- tolyl) -1H - pyrazolo [3,4- c ] pyridin-7 ( 4H ) -one in the same manner as in Example 1 above (94 mg, 0.388 mmol), potassium carbonate (80.4 mg, 0.582 mmol) and 1- {4- (4 -t- butylphenyl) piperazin-1-yl} -3-chloropropan-1-one (131.5 mg , 0.426 mmol) to give 173.3 mg (0.336 mmol, 86.8%) of the title compound in the solid state.

¹ H NMR (CDCl ₃ , 300 MHz) δ 7.28 (d, J = 8.7 Hz, 2H), 7.19-7.12 (m, 4H), 6.83 (d, J = 8.7 Hz, 2H), 4.53 (s, 2H, NH ₂ ), 4.36 (t, J = 4.8 Hz, 2H), 3.82 (t, J = 6.4 Hz, 2H), 3.68 (br, 2H), 3.54 (br, 2H), 3.04 (br, 4H), 2.97 (t, J = 4.8 Hz, 2H), 2.65 (t, J = 6.4 Hz, 2H), 2.31 (s, 3H), 1.28 (s, 9H)

¹³ C NMR (CDCl ₃ , 75 MHz) δ 169.6, 161.6, 148.4, 143.3, 142.2, 142.0, 140.5, 135.7, 129.4, 126.0, 125.5, 116.3, 102.2, 51.9, 49.6, 49.4, 45.3, 43.0, 41.8, 34.0 , 33.3, 31.4, 21.0, 19.5

Example 28. 3-Amino-1-[{4- (cyclohexylphenyl) piperazin-1-yl} propanoyl] -6- N- ( p -tolyl) -4,5,6,7-tetra Hydro-1 H -pyrazolo [3,4- c ] pyridin-7-one (Compound No. 28)

3-Amino-5,6-dihydro-6- N- ( p- tolyl) -1H - pyrazolo [3,4- c ] pyridin-7 ( 4H ) -one in the same manner as in Example 1 above (90 mg, 0.371 mmol), potassium carbonate (76.8 mg, 0.556 mmol) and 3-chloro-1- {4- (4-cyclohexylphenyl) piperazin-1-yl} propan-1-one (136.6 mg, 0.408 mmol) was reacted to give 136.1 mg (0.251 mmol, 67.8%) of the title compound in the solid state.

¹ H NMR (CDCl ₃ , 300 MHz) δ 7.19-7.15 (m, 4H), 7.10 (d, J = 8.6 Hz, 2H), 6.82 (d, J = 8.5 Hz, 2H), 4.51 (s, 2H, NH ₂ ), 4.35 (t, J = 4.7 Hz, 2H), 3.82 (t, J = 6.3 Hz, 2H), 3.70 (br, 2H), 3.54 (br, 2H), 3.03 (br, 4H), 2.97 (t, J = 4.8 Hz, 2H), 2.66 (t, J = 6.3 Hz, 2H), 2.42 (br, 1H), 2.31 (s, 3H), 1.81 (br, 4H), 1.73 (d, J = 12.2 Hz, 1H), 1.43-1.20 (m, 5H)

¹³ C NMR (CDCl ₃ , 75 MHz) δ 169.6, 161.6, 148.8, 142.2, 141.9, 140.5, 140.4, 135.7, 129.4, 127.4, 125.5, 116.7, 102.2, 51.9, 49.8, 49.6, 45.4, 43.6, 42.9, 41.9 , 34.6, 33.2, 26.9, 26.1, 21.0, 19.4

Formulation Example

On the other hand, the novel compound represented by Formula 1 according to the present invention can be formulated in various forms according to the purpose. The following is a description of some formulations containing the compound of Formula 1 according to the present invention as an active ingredient, but the present invention is not limited thereto.

Formulation 1: tablets (direct pressure)

After 5.0 mg of the active ingredient was sieved, 14.1 mg of lactose, 0.8 mg of crospovidone USNF and 0.1 mg of magnesium stearate were mixed and pressurized to make tablets.

Formulation 2: tablets (wet granulation)

After 5.0 mg of the active ingredient was sieved, 16.0 mg of lactose and 4.0 mg of starch were mixed. 0.3 mg of Polysorbate 80 was dissolved in pure water, and an appropriate amount of this solution was added, followed by atomization. After drying, the fine particles were sieved and mixed with 2.7 mg of colloidal silicon dioxide and 2.0 mg of magnesium stearate. The granules were pressed into tablets.

Formulation 3: Powders and Capsules

5.0 mg of the active ingredient was sieved, followed by mixing with 14.8 mg of lactose, 10.0 mg of polyvinyl pyrrolidone, and 0.2 mg of magnesium stearate. The mixture was filtered through a hard No. Filled in 5 gelatin capsules.

Formulation 4: Injection

Injectables were prepared by containing 100 mg as active ingredient, as well as 180 mg of mannitol, 26 mg of Na ₂ HPO ₄ 12H ₂ O and 2974 mg of distilled water.

Experimental Example

Anticancer Drug Test

1) Cancer Cell Culture

Cancer cells to measure anticancer activity against test compounds A-549 (Human lung carcinoma), HT-29 (Human colon adenocarcinoma), DU145 (Human prostate cancer), SK-MEL-2 (Human malignant melanoma), and SK-OV-3 (Human ovary malignant ascites) were used. . These cancer cells were all human tumor cells and were cultured by the Korean Cell Line Bank. Culture medium was used RP MI 1640 medium containing 10% fetal calf serum, and cultured in a constant temperature and humidity incubator (37 ℃, 5% CO ₂₎ . Cell passage was performed once every 3 days using 0.25% trypsin - 1 mM EDTA.

2) Anticancer activity measurement

In 1989, the US National Cancer Institute used the SRB assay (Sulforhodamine B assay method) developed to measure the in vitro anticancer activity of the drug.

In order to use the passaged cells for experiments, cells were separated from the implant using trypsin-EDTA solution, and the cells in culture were dispensed in 96 well microplates so that the number of cells per well was 5 × 10 ³ for 24 hours in a CO ₂ incubator. Incubated. The medium was removed, and 100 μl of the test compound solution diluted to 4-fold concentration was incubated for 48 hours. 100 μl of formalin solution was added to fix the cells, and then washed five times with distilled water and dried at room temperature. 100 μl of 0.4% SRB solution was added thereto, and the mixture was left at room temperature for 30 minutes, washed five times with 1% acetic acid, and dried at room temperature. 200 μl of 10 mM Tisma base pH 10.3 was added to each well to completely dissolve the absorbance at 520 nm.

In order to calculate the efficacy of the test compound on cancer cells, GI ₅₀ was calculated according to Equation 1 or 2 below.

In Equation 1, T ₀ is the number of cells before adding the test compound, and T ₂ is the number of cells after incubation for 48 hours after adding the test compound.

In Equation 2, T ₀ is the number of cells before adding the test compound, T ₂ is the number of cells after incubation for 48 hours after adding the test compound, and C is 48 hours incubating the control group without adding the test compound. The number of cells after one.

In addition, by using the data regression by the Lotus program (LOTUS program) from the values calculated by the equation (1) to obtain the degree (% inhibition concentration) to inhibit the growth of each test compound cancer cells Table 1 Shown in

Experimental compound % Inhibition rate for cancer cell lines (100 mM) DU-145 HT-29 HCT-116 A375P Compound number 2 75.05 95.21 72.18 86.03 Compound number 3 21.27 38.19 9.95 38.1 Compound number 4 90.55 94.07 93.17 89.73 Compound No. 7 95.97 96.75 97.39 97.62 Compound number 8 52.38 45.06 42.06 43.38 Compound number 9 80.02 90.54 92.19 95.03 Compound number 10 38.10 32.12 33.48 44.03 Compound number 12 85.77 90.68 96.57 97.26 Compound number 13 24.99 29.50 36.37 22.49 Compound number 15 72.19 74.12 79.42 89.14 Compound number 17 93.50 94.43 95.46 97.28 Doxorubicin 62.79 60.89 N.D. N.D. DU-145: human prostate cancer,
HT-29: human colon adenocarcinoma cell,
HCT-116: human colon cancer,
A375P: human melanoma cancer

As confirmed in Table 1, 1,6-disubstituted-3-amino-4,5,6,7-tetrahydro- 1H -pyrazolo [3,4- c ] represented by Chemical Formula 1 according to the present invention. Pyridin-7-one compounds are cytotoxic to various cancer cells and are therefore useful as new anticancer agents.

Claims (6)

1,6-disubstituted-3-amino-4,5,6,7-tetrahydro- 1H -pyrazolo [3,4- c ] pyridin-7-one compound represented by Formula 1 below or a pharmaceutically acceptable Compounds selected from salts thereof:
[Formula 1]

In Chemical Formula 1,
n represents an integer of 1, 2, or 3,
R ¹ represents R— (CH ₂ ) _L −, where R is substituted with one to three substituents selected from halo, C ₁ -C ₈ alkyl, haloC ₁ -C ₈ alkyl, and C ₁ -C ₈ alkoxy Or an unsubstituted phenyl group; Furyl group; Pyridyl groups; Or a benzodioxol group,
R ² represents R— (CH ₂ ) _L −, where R is C ₁ -C ₈ alkyl, C ₁ -C ₈ alkoxy, amino, C ₁ -C ₈ alkylamino, and di (C ₁ -C ₈ alkyl Phenyl group unsubstituted or substituted with 1 to 3 substituents selected from amino; Or a furyl group,
L represents an integer of 0, 1, 2 or 3.
The method according to claim 1,
N represents an integer of 1, 2, or 3,
R ¹ is a phenyl group, 3-chlorophenyl group, 4-chlorophenyl group, 2,3-dichlorophenyl group, 2,3-difluorophenyl group, 4- (trifulomethyl) phenyl group, 2-methylphenyl group, 3 -Methylphenyl group, 4-methylphenyl group, 4- t -butylphenyl group, 4-cyclohexylphenyl group, 2,3-dimethylphenyl group, 2,4-dimethylphenyl group, 2,4,5-trimethylphenyl group, 4-meth Methoxyphenyl group, benzyl group, 4- t -butylbenzyl group, 2,4,6-trimethylbenzyl group, 2,3,4-trimethoxybenzyl group, (benzo [ d ] [1,3] dioxol-5 -Yl) methyl group, furyl group, furan-2-ylmethyl group, or pyridin-2-yl group,
R ² is a phenyl group, 2-methylphenyl group, 3-methylphenyl group, 4-methylphenyl group, 4- t -butylphenyl group, 4-cyclohexylphenyl group, 2,3-dimethylphenyl group, 2,4-dimethylphenyl group, 2,4,5-trimethylphenyl group, 4-methoxyphenyl group, 4-dimethylaminophenyl group, benzyl group, 4- t -butylbenzyl group, 2,4,6-trimethylbenzyl group, 2,3,4-tri A methoxy benzyl group, a furyl group, or a furan-2-ylmethyl group.
The method according to claim 1,
3-amino-1-[{4- (2,3-dimethylphenyl) piperazin-1-yl} ethanoyl] -6- N- ( p- tolyl) -4,5,6,7-tetrahydro- 1 H- pyrazolo [3,4- c ] pyridin-7-one (Compound No. 1),
3-amino-1-[{4- (2,3-dimethylphenyl) piperazin-1-yl} propanoyl] -6- N- ( p- tolyl) -4,5,6,7-tetrahydro -1 H- pyrazolo [3,4- c ] pyridin-7-one (Compound No. 2),
3-amino-1-{(4-phenylpiperazin-1-yl) propanoyl} -6- N- ( p- tolyl) -4,5,6,7-tetrahydro-1 H- pyrazolo [ 3,4- c ] pyridin-7-one (Compound No. 3),
3-amino-1-[{4- (2,4-dimethylphenyl) piperazin-1-yl} propanoyl] -6- N- ( p- tolyl) -4,5,6,7-tetrahydro -1 H- pyrazolo [3,4- c ] pyridin-7-one (Compound No. 4),
3-amino-1-[{4-((benzo [ d ] [1,3] dioxol-5-yl) methyl) piperazin-1-yl} propanoyl] -6- N- ( p- tolyl ) -4,5,6,7-tetrahydro-1 H- pyrazolo [3,4- c ] pyridin-7-one (Compound No. 5),
3-amino-1-[{4- (2,4,6-trimethylbenzyl) piperazin-1-yl} propanoyl] -6- N- ( p- tolyl) -4,5,6,7- Tetrahydro-1 H- pyrazolo [3,4- c ] pyridin-7-one (Compound No. 6),
3-amino-1-[{4- (4- t- butylbenzyl) piperazin-1-yl} propanoyl] -6- N- ( p- tolyl) -4,5,6,7-tetrahydro -1 H- pyrazolo [3,4- c ] pyridin-7-one (Compound No. 7),
3-amino-1-[{4- (2,3,4-trimethoxybenzyl) piperazin-1-yl} propanoyl] -6- N- ( p- tolyl) -4,5,6, 7-tetrahydro-1 H- pyrazolo [3,4- c ] pyridin-7-one (Compound No. 8),
3-amino-1-[{4- (4-chlorophenyl) piperazin-1-yl} propanoyl] -6- N- ( p- tolyl) -4,5,6,7-tetrahydro-1 H- pyrazolo [3,4- c ] pyridin-7-one (Compound No. 9),
3-amino-1-[{4- (4-methoxyphenyl) piperazin-1-yl} propanoyl] -6- N- ( p- tolyl) -4,5,6,7-tetrahydro- 1 H- pyrazolo [3,4- c ] pyridin-7-one (Compound No. 10),
3-amino -1 - {(N-benzyl) piperazin-1-yl} -6- N - (p- methoxyphenyl) 4,5,6,7-tetrahydro-pyrazolo [3,4 - c ] pyridin-7-one (Compound No. 11),
3-amino-1-[{4- (2,4-dimethylphenyl) piperazin-1-yl} propanoyl] -6- N- ( p -methoxyphenyl) -4,5,6,7- Tetrahydro-1 H- pyrazolo [3,4- c ] pyridin-7-one (Compound No. 12),
3-amino-1-[{4- (2,4,6-trimethylbenzyl) piperazin-1-yl} propanoyl] -6- N- ( p -methoxyphenyl) -4,5,6, 7-tetrahydro-1 H- pyrazolo [3,4- c ] pyridin-7-one (Compound No. 13),
3-amino-1-[{4- (2,3,4-trimethoxybenzyl) piperazin-1-yl} propanoyl] -6- N- ( p -methoxyphenyl) -4,5, 6,7-tetrahydro-1 H- pyrazolo [3,4- c ] pyridin-7-one (Compound No. 14),
3-amino-1-[{4- (2,4-dimethylphenyl) piperazin-1-yl} propanoyl] -6- N -phenyl-4,5,6,7-tetrahydro-1 H- Pyrazolo [3,4- c ] pyridin-7-one (Compound No. 15),
3-amino-1-[{4- (2,4,6-trimethylbenzyl) piperazin-1-yl} propanoyl] -6- N -phenyl-4,5,6,7-tetrahydro-1 H- pyrazolo [3,4- c ] pyridin-7-one (Compound No. 16),
3-amino-1-[{4- (2,4-dimethylphenyl) piperazin-1-yl} propanoyl] -6- N -benzyl-4,5,6,7-tetrahydro-1 H- Pyrazolo [3,4- c ] pyridin-7-one (Compound No. 17),
3-amino-1-{(4-phenylpiperazin-1-yl) propanoyl} -6- N- { p- (dimethylamino) phenyl} -4,5,6,7-tetrahydro-1 H- pyrazolo [3,4- c ] pyridin-7-one (Compound No. 18),
3-amino-1-[{4- (4-methoxyphenyl) piperazin-1-yl} propanoyl] -6- N- { p- (dimethylamino) phenyl} -4,5,6, 7-tetrahydro-1 H- pyrazolo [3,4- c ] pyridin-7-one (Compound No. 19),
3-amino-1-{(4-phenylpiperazin-1-yl) propanoyl} -6- N -{(furan-2-yl) methyl} -4,5,6,7-tetrahydro-1 H- pyrazolo [3,4- c ] pyridin-7-one (Compound No. 20),
3-amino-1-[{4- (4-methoxyphenyl) piperazin-1-yl} propanoyl] -6- N -{(furan-2-yl) methyl} -4,5,6, 7-tetrahydro-1 H- pyrazolo [3,4- c ] pyridin-7-one (Compound No. 21),
3-amino-1- {4- (2-pyridinylpiperazin-1-yl) propanoyl} -6- Np- tolyl-4,5,6,7-tetrahydro-1 H -pyrazolo [3 , 4- c ] pyridin-7-one (Compound No. 22),
3-amino-1- {4- (2-pyridinylpiperazin-1-yl) propanoyl} -6- N -{(furan-2-yl) methyl} -4,5,6,7-tetra Hydro-1 H -pyrazolo [3,4- c ] pyridin-7-one (Compound No. 23),
3-amino-1-[{4- (4-trifluorofluoromethylphenyl) piperazin-1-yl} propanoyl] -6- N- ( p- tolyl) -4,5,6,7-tetra Hydro-1 H -pyrazolo [3,4- c ] pyridin-7-one (Compound No. 24),
3-amino-1-[{4- (2,4-dimethylphenyl) piperazin-1-yl} propanoyl] -6- N -{(furan-2-yl) methyl} -4,5,6 , 7-tetrahydro-1 H -pyrazolo [3,4- c ] pyridin-7-one (Compound No. 25),
3-amino-1-[{4- (2,4,5-trimethylphenyl) piperazin-1-yl} propanoyl] -6- N- ( p- tolyl) -4,5,6,7- Tetrahydro-1 H -pyrazolo [3,4- c ] pyridin-7-one (Compound No. 26),
3-amino-1-[{4- (4- t- butylphenyl) piperazin-1-yl} propanoyl] -6- N- ( p- tolyl) -4,5,6,7-tetrahydro -1 H -pyrazolo [3,4- c ] pyridin-7-one (Compound No. 27),
3-amino-1-[{4- (cyclohexylphenyl) piperazin-1-yl} propanoyl] -6- N- ( p- tolyl) -4,5,6,7-tetrahydro-1 H -Pyrazolo [3,4- c ] pyridin-7-one (Compound No. 28), and
Compound selected from pharmaceutically acceptable salts thereof.
A pharmaceutical composition for anticancer drugs, characterized in that the compound of any one selected from claims 1 to 3 contained as an active ingredient.
1,6-disubstituted-3, wherein the 3-amino compound represented by the following Formula 2 and the haloalkylcarbonyl peparagine compound represented by the following Formula 3 are alkylated to prepare a compound represented by the following Formula 1. Method for preparing -amino-4,5,6,7-tetrahydro- 1H -pyrazolo [3,4- c ] pyridin-7-one compound:

In the above scheme, n, R ¹ , and R ² are each as defined in claim 1, and X represents a halogen atom.
3-Amino compound represented by Formula 2 below:
(2)

In Formula 2, R ¹ is as defined in claim 1 above.